Compositions

ABSTRACT

There is provided a solid pharmaceutical composition for delivering by oral administration a pharmaceutically active binding polypeptide to a region of the intestinal tract comprising a compressed core, wherein the compressed core comprises a pharmaceutically active binding polypeptide and wherein the compressed core is coated with a pH sensitive enteric coating.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/140,843, filed on Sep. 25, 2018, which is a continuation ofInternational Application No. PCT/EP2017/057775, filed on Mar. 31, 2017,which claims priority to European Application No. 16163178.3, filed onMar. 31, 2016, the contents of each of which are herein incorporated byreference in their entirety.

FIELD OF THE INVENTION

The present invention relates to solid pharmaceutical compositions forthe purpose of delivering by oral administration a pharmaceuticallyactive binding polypeptide to a region of the intestinal tract. Thesecompositions benefit from advantageous release profiles and may be usedin the treatment of diseases of the intestinal tract. The presentinvention also relates to methods of delivering pharmaceutically activebinding polypeptides to a region of the intestinal tract by oraladministration.

BACKGROUND OF THE INVENTION

Diseases, such as inflammatory bowel disease, manifest in variousregions of the intestinal tract, such as regions of the small intestine.There exist pharmaceutically active binding polypeptides which areeffective in the treatment of inflammatory bowel disease whenadministered systemically. For example, anti-TNF-alpha antibodies havedemonstrated efficacy in treating inflammatory bowel diseases. However,because these antibodies are commonly delivered by injection (i.e.intravenous, subcutaneous or intramuscular) and neutralise TNF-alphasystemically, their use may be associated with serious side effects,including reactivation of tuberculosis and a long-term risk ofmalignancy. Moreover, the parenteral route of administration and thelarge doses required make these antibody therapies expensive and hardlyaccessible for patients.

Oral administration of such polypeptides for local effect in a targetregion of the intestinal tract would be preferable, due to for examplereduced cost and the convenience of this dosage form. In addition, oraladministration may provide reduced immunogenicity as compared toparenteral administration forms and may reduce or eliminate unnecessarysystemic exposure to the polypeptide.

To achieve this goal, a suitable release profile must be achieved. Thatis, the activity of the polypeptide must be maintained after transitthrough the upper intestinal tract including the stomach and suitablythe duodenum and the desired dosage of active polypeptide must bedelivered to the desired location of the intestinal tract.

Pharmaceutical compositions of the present invention may, in at leastsome embodiments, have one or more of the following advantages comparedto those of the prior art:

-   -   (i) a sustained release profile,    -   (ii) a delayed release profile,    -   (iii) targeted release to one or more regions of the intestinal        tract,    -   (iv) substantially consistent release to all regions of the        intestinal tract from the duodenum to the anal canal (i.e.        avoiding ‘dose dumping’),    -   (v) reduced host immune response to the delivered polypeptide        compared to parenteral administration,    -   (vi) reduced systemic exposure to pharmaceutically active agent,    -   (vii) reduced dosage required for therapeutic effects,    -   (viii) reduced cost of production,    -   (ix) maintained or improved thermal stability of polypeptide.

PRIOR ART

WO2014/030049 (D1) discloses compositions comprising a single variabledomain and camostat mesylate as a means of stabilising a single variabledomain, in particular in protease-rich environments such as the stomachand intestine. D1 exemplifies liquid compositions, which are injecteddirectly into the gastrointestinal tract of mice.

US2010/260857 (D2) discloses coated digestive enzyme preparations. D2exemplifies free-flowing digestive enzyme particles coated with lipidsand packaged in a sachet or capsule.

WO2008/122965 (D3) discloses cyclosporin compositions in a solubilisedliquid form.

US2006/057197 (D4) discloses pharmaceutical dosage forms fornon-polypeptide, small molecules. All exemplification in D4 relates tothe delivery of baclofen and subsequent monitoring of plasma profiles.

Hussan et al 2012 IOSR Journal of Pharmacy 2(6):5-11 (D5) is a review ofrecent advances in enteric coating.

Harmsen et al 2006 Applied Microbiology and Biotechnology 72(3):544-551(D6) discloses the selection and optimization of proteolytically stablellama single-domain antibody fragments for oral immunotherapy. D6provides no information on solid dosage forms.

Hussack et al 2011 PLOS ONE 6(11):e28218 (D7) discloses engineeredsingle-domain antibodies with high protease resistance and thermalstability. D7 provides no information on solid dosage forms.

None of the above discloses a solid composition comprising a compressedcore capable of sustained intestinal delivery of pharmaceutically activebinding polypeptide.

SUMMARY OF THE INVENTION

The present inventors have produced surprisingly advantageous solidpharmaceutical formulations suitable for delivering by oraladministration a pharmaceutically active binding polypeptide to a regionof the intestinal tract. These pharmaceutical formulations areparticularly advantageous due to their delayed and/or sustained releaseprofiles. It may be expected that these pharmaceutical formulations haveparticular utility in the prevention or treatment of diseases of theintestinal tract such as autoimmune and/or inflammatory disease such asinflammatory bowel disease, or in the prevention or treatment ofinfection from an intestinal tract resident pathogenic microbe.

The present invention provides a solid pharmaceutical composition fordelivering by oral administration a pharmaceutically active bindingpolypeptide to a region of the intestinal tract comprising a compressedcore, wherein the compressed core comprises a pharmaceutically activebinding polypeptide and wherein the compressed core is coated with a pHsensitive enteric coating.

Also provided is a solid pharmaceutical composition for use in thetreatment of a disease of the intestinal tract by oral administration toa region of the intestinal tract comprising a compressed core, whereinthe compressed core comprises a pharmaceutically active bindingpolypeptide and wherein the compressed core is coated with a pHsensitive enteric coating.

Also provided is a method of delivering a pharmaceutically activebinding polypeptide to a region of the intestinal tract comprisingorally administering a solid pharmaceutical composition comprising acompressed core, wherein the compressed core comprises apharmaceutically active binding polypeptide and wherein the compressedcore is coated with a pH sensitive enteric coating.

Also provided is a compressed core for a solid pharmaceuticalcomposition wherein the composition is for delivering a pharmaceuticallyactive binding polypeptide to a region of the intestinal tract by oraladministration and wherein the compressed core comprises apharmaceutically active binding polypeptide for local therapeuticeffect.

DESCRIPTION OF THE FIGURES

FIG. 1—% dissolution of pharmaceutically active binding polypeptide inThe Pharmacopeial Dissolution Test (individual data points)

FIG. 2—% dissolution of pharmaceutically active binding polypeptide inThe Pharmacopeial Dissolution Test (average of data points)

FIG. 3—Simulated stomach and intestinal pH profile for fastingconditions used in The Dynamic Dissolution Test

FIG. 4—% dissolution of pharmaceutically active binding polypeptide inThe Dynamic Dissolution Test (Batch A)

FIG. 5—% dissolution of pharmaceutically active binding polypeptide inThe Dynamic Dissolution Test (Batch B)

FIG. 6—Thermal shift assay demonstrating the impact of excipients on Tmof pharmaceutically active binding polypeptide

FIG. 7—Calculated luminal [anti-TNF ICVD] in cynomolgus monkeygastrointestinal tract sections

FIG. 8—Total % recovery of anti-TNF ICVD from cynomolgus monkeygastrointestinal tracts

FIG. 9—Humira competition ELISA OD450 data

FIG. 10—anti-TNF ICVD concentration in pooled cynomolgus monkey faeces

FIG. 11—Calculated anti-TNF ICVD recovered from pooled cynomolgus monkeyfaeces

FIG. 12—Dynamic Dissolution Test comparing spray dried and lyophilisedstarting materials

DESCRIPTION OF THE SEQUENCES

-   SEQ ID NO: 1—Polypeptide sequence of Q62E10-   SEQ ID NO: 2—Polypeptide sequence of Q65F2-   SEQ ID NO: 3—Polypeptide sequence of Q65F3-   SEQ ID NO: 4—Polypeptide sequence of Q62F2-   SEQ ID NO: 5—Polypeptide sequence of Q65G1-   SEQ ID NO: 6—Polypeptide sequence of Q65H6-   SEQ ID NO: 7—Polypeptide sequence of Q65F1-   SEQ ID NO: 8—Polypeptide sequence of Q65D1-   SEQ ID NO: 9—Polypeptide sequence of Q65C7-   SEQ ID NO: 10—Polypeptide sequence of Q65D3-   SEQ ID NO: 11—Polypeptide sequence of Q65B1-   SEQ ID NO: 12—Polypeptide sequence of Q65F6-   SEQ ID NO: 13—Polypeptide sequence of Q65F11-   SEQ ID NO: 14—Polypeptide sequence of Q65E12-   SEQ ID NO: 15—Polypeptide sequence of Q65C12-   SEQ ID NO: 16—Polypeptide sequence of Q65A6-   SEQ ID NO: 17—Polypeptide sequence of Q65A3-   SEQ ID NO: 18—Polypeptide sequence of Q62F10-   SEQ ID NO: 19—Polypeptide sequence of ID7F-EV-   SEQ ID NO: 20—Polypeptide sequence of ID8F-EV-   SEQ ID NO: 21—Polypeptide sequence of ID9F-EV-   SEQ ID NO: 22—Polypeptide sequence of ID13F-EV-   SEQ ID NO: 23—Polypeptide sequence of ID14F-EV-   SEQ ID NO: 24—Polypeptide sequence of ID15F-EV-   SEQ ID NO: 25—Polypeptide sequence of Q62E10-DVQLV-   SEQ ID NO: 26—Polypeptide sequence of ID34F-   SEQ ID NO: 27—Polypeptide sequence of ID37F-   SEQ ID NO: 28—Polypeptide sequence of ID38F

DETAILED DESCRIPTION OF THE INVENTION

Release Profile

The release profile of a composition comprising a pharmaceuticallyactive binding polypeptide is the quantity of pharmaceutically activebinding polypeptide released from the composition, and is therefore freeto bind its target, over time. The present application is concerned withthe release profile of solid pharmaceutical compositions in theintestinal tract from the stomach to the rectum. Release profile mayrefer to that which is achieved in vivo during transit from the stomachto the rectum, or that which is achieved in an in vitro model of transitfrom the stomach to the rectum. Release profiles may be tested in vitroby dissolution testing using suitable dissolution apparatus, such asthose described below. More suitably, the present application isconcerned with the release profile of solid pharmaceutical compositionsin the intestinal tract from the duodenum to the rectum and moresuitably from the jejunum to the rectum.

The European Medicines Agency's “Guideline on quality of oral modifiedrelease products” under section “3.2. Setting specifications” statesthat at least two points should be included in a specification on invitro dissolution of a gastro-resistant product: an early time point toexclude release in the acidic medium (less than 10% dissolved after 2hours) and one to ensure that the majority of the active substance hasbeen released in a (near) neutral medium.

“Delayed release” refers to the ability of a composition comprising apharmaceutically active binding polypeptide to (a) protect thepharmaceutically active binding polypeptide from the externalenvironment (e.g. acidic pH) and (b) not release any pharmaceuticallyactive binding polypeptide to the external environment, until thecomposition reaches a desired region of the intestinal tract. Delayedrelease is achieved by virtue of a pH sensitive enteric coating. Thelonger the pH sensitive enteric coating maintains integrity, the longerthe period of delayed release. The properties of the pH sensitiveenteric coating may be adapted such that release may be additionallydelayed (for example to ensure pharmaceutically active bindingpolypeptide is not released until the composition has passed theduodenum and has entered the jejunum). Alternatively, the properties ofthe pH sensitive enteric coating may be adapted such that release may beadvanced (for example to ensure pharmaceutically active bindingpolypeptide is released only after exiting the stomach but while stillinside the duodenum).

“Sustained release” refers to the ability of a composition comprising apharmaceutically active binding polypeptide to release pharmaceuticallyactive binding polypeptide into the external environment at a desired,ideally substantially constant rate, during transit through desiredregions (or a desired region) of the intestinal tract.

A composition of the invention will suitably achieve both delayedrelease and sustained release.

There exist official in vitro tests of delayed and sustained releasewhich are comprehensively defined in Pharmacopoeia. Tablet dissolutionis a standardised method for measuring the rate of drug release from adosage form. In vitro dissolution tests can be used to predict in vivodrug dissolution. Suitable in vitro dissolution tests are detailedbelow.

Delayed Release: Enteric Coating Release Profile

Measurement of Delayed Release: The Pharmacopeial Enteric Coating Test

A suitable and widely used standard method of testing an enteric coatingis that provided by the European Pharmacopoeia 8.0 “2.9.3 Dissolutiontest for solid dosage forms” (which is harmonised with the correspondingtexts of the United States Pharmacopeia and the Japanese Pharmacopoeia),referred to herein as the “Pharmacopeial Enteric Coating Test”. Thisenteric coating test is carried out as follows.

1. The composition to be tested is added to 900 mL of 0.1M HCl in USP2apparatus (with band sinkers if presented in a capsule).

2. The composition and acid is stirred at 100 rpm for 2 hours.

The dissolution medium is sampled and analysed at 2 hours. Suitably thepharmaceutical composition releases less than 10% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases less than 5% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases less than 1% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases no pharmaceutically activebinding polypeptide after 2 hours.

Release of less than 10% by weight of the pharmaceutically activebinding polypeptide after 2 hours indicates that the enteric coatingprovides adequate protection to the polypeptide preceding delivery tothe desired region of the intestinal tract. A composition with thisrelease profile achieves delayed release.

If desired, The Pharmacopeial Enteric Coating Test may be modified tocontinue to run for longer than 2 hours. This will allow analysis of themaximum time over which the pH sensitive enteric coating remains intact.When using small quantities of composition, accuracy of the test may beincreased by reducing the volume of HCl used.

A pH sensitive enteric coating is said to be ‘intact’ when, based onvisual inspection, the composition shows no signs of eitherdisintegration or cracks that would allow the escape of the contents (asoutlined in European Pharmacopoeia 8.0 2.9.3). Alternatively, a pHsensitive enteric coating can be said to be ‘intact’ when 1% or less,more suitably 0.5% or less or more suitably no pharmaceutically activebinding polypeptide can be detected in the dissolution medium in whichthe composition is present. Suitably, the pH sensitive enteric coatingremains intact for at least 2 hours, more suitably at least 3 hours,more suitably at least 4 hours, more suitably at least 5 hours, moresuitably at least 6 hours during the Pharmacopeial Enteric Coating Test.

Sustained Release: Core Release Profile

Sustained release properties are suitably tested immediately after acomposition has undergone testing for delayed release properties usingThe Pharmacopeial Enteric Coating Test detailed above. If this is thecase, the composition should be sieved from the 0.1M HCl after delayedrelease test completion and washed with fresh 0.1M HCl, before finallytransferring the composition to the dissolution media (phosphate buffer)described in the sustained release test described below.

Measurement of sustained release: The Pharmacopeial Dissolution Test

A suitable and widely used standard method of testing dissolution isthat provided by the European Pharmacopoeia 8.0 “2.9.3 Dissolution testfor solid dosage forms” (which is harmonised with the correspondingtexts of the United States Pharmacopeia and the Japanese Pharmacopoeia).This dissolution test is referred to herein as the “PharmacopeialDissolution Test”. This dissolution test is carried out as follows.

1. The composition to be tested is added to 900 mL of 0.05M pH 7.4phosphate buffer in USP2 apparatus (with band sinkers if presented in acapsule).

2. The composition and buffer is stirred at 100 rpm for 2 hours.

3. The composition and buffer is then stirred at 200 rpm for 30 mins.

The dissolution medium may be sampled and analysed at 5, 10 15, 30, 45,60, 90 and 120 minutes (during the 100 rpm stirring period) and at 150minutes (during the 200 rpm stirring period).

Suitably, when assayed in the Pharmacopeial Dissolution Test, thepharmaceutical composition of the invention releases:

(i) 10-40% by weight of the pharmaceutically active binding polypeptideafter 30 minutes,

(ii) 30-60% by weight of the pharmaceutically active binding polypeptideafter 60 minutes and

(iii) 60% by weight or greater of the pharmaceutically active bindingpolypeptide after 120 minutes.

FIG. 2 illustrates the % dissolution profile achieved by the compositionof Example 1 in the Pharmacopeial Dissolution Test. The dissolutionranges above are illustrated by arrows in FIG. 2.

More suitably, when assayed in the Pharmacopeial Dissolution Test, thepharmaceutical composition of the invention releases:

(i) 10-25% by weight of the pharmaceutically active binding polypeptideafter 30 minutes,

(ii) 30-50% by weight of the pharmaceutically active binding polypeptideafter 60 minutes and

(iii) 60% by weight or greater of the pharmaceutically active bindingpolypeptide after 120 minutes.

More suitably, when assayed in the Pharmacopeial Dissolution Test, thepharmaceutical composition of the invention releases:

(i) 13-20% by weight of the pharmaceutically active binding polypeptideafter 30 minutes,

(ii) 35-45% by weight of the pharmaceutically active binding polypeptideafter 60 minutes and

(iii) 65% by weight or greater of the pharmaceutically active bindingpolypeptide after 120 minutes.

Delayed and Sustained Release: Joint Enteric Coating and Core ReleaseProfile

The Dynamic Dissolution Test

Alternatively, or in addition to the Pharmacopeial tests describedabove, a suitable method of jointly testing an enteric coating and corerelease profile is referred to herein as “The Dynamic Dissolution Test”.

FIG. 3 illustrates the simulated pH profile for fasting conditions whichis used in The Dynamic Dissolution Test. Further information on thistest is available primarily in Garbacz et al 2014 and also in Fadda etal 2009, Merchant et al 2014, and Goyanes et al 2015. This test iscarried out as follows.

1. The composition to be tested is added to 900 mL of 0.1M HCl in USP2apparatus.

2. The composition and acid is stirred at 50 rpm for 2 hours.

This 2 hour period in 0.1M HCl is denoted by the region of FIG. 3preceding the dotted line.

The dissolution medium is sampled and analysed at 2 hours. Suitably thepharmaceutical composition releases less than 10% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases less than 5% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases less than 1% by weight of thepharmaceutically active binding polypeptide after 2 hours. More suitablythe pharmaceutical composition releases no pharmaceutically activebinding polypeptide after 2 hours.

Release of less than 10% by weight of the pharmaceutically activebinding polypeptide after 2 hours indicates that the enteric coatingprovides adequate protection to the polypeptide preceding delivery tothe desired region of the intestinal tract. A composition with thisrelease profile achieves delayed release.

The next stage of this test ascertains the dissolution of a compositionduring transit through the varying pH profile existent along thegastrointestinal tract. FIG. 3 (curve following the dotted line)illustrates the simulated pH profile for fasting conditions which isused. This stage of the test is carried out as follows.

1. The composition being tested is transferred to 900 mL of pH 5.79Hanks hydrogen carbonate buffer (details in Table 1 below) in USP2apparatus (the moment of transfer to Hanks buffer is denoted by thedotted line in FIG. 3).

2. The composition and buffer is stirred at 50 rpm for 16 hours overwhich time the pH of the dissolution medium is varied according to FIG.3. Variation of the dissolution medium pH may be varied by exposure togaseous carbon dioxide (to lower pH) or gaseous nitrogen (to raise pH).The pH profile simulates a median pH profile of small bowel passage aswell as model pH profile of colonic transit, during fasting.

3. The composition and buffer is then stirred at 200 rpm for 2 hours.

TABLE 1 Amount for 1 L (g) Compound CAS Number 0.06 Potassium dihydrogenphosphate 7778-77-0 8.00 Sodium chloride 7647-14-5 0.03 Sodiumdihydrogen phosphate dihydrate 13472-35-0 0.04 Calcium chloride10035-04-8 0.40 Potassium chloride 7447-40-7 0.20 Magnesium sulfate10034-99-8 0.35 Sodium hydrogen carbonate 144-55-8

Samples may be taken every 10 minutes for the duration of the test foranalysis of the quantity of released pharmaceutically active bindingpolypeptide.

Suitably the start of release of the pharmaceutically active bindingpolypeptide occurs between 90 to 210 minutes from addition of thecomposition to Hanks buffer. ‘Start of release’ as used herein refers tothe first point in the Dynamic Dissolution Test after addition to Hanksbuffer at which greater than 1% of pharmaceutically active bindingpolypeptide is released from the core.

Suitably, when assayed in the Dynamic Dissolution Test, thepharmaceutical composition releases:

(i) 10-30% by weight of the pharmaceutically active binding polypeptideafter 60 minutes from start of release,

(ii) 40-70% by weight of the pharmaceutically active binding polypeptideafter 120 minutes from start of release and

(iii) 60% by weight or greater of the pharmaceutically active bindingpolypeptide after 180 minutes from start of release.

Suitably, when assayed in the Dynamic Dissolution Test, thepharmaceutical composition releases:

(i) 10-30% by weight of the pharmaceutically active binding polypeptideafter 60 minutes from start of release,

(ii) 40-60% by weight of the pharmaceutically active binding polypeptideafter 120 minutes from start of release and

(iii) 60% by weight or greater of the pharmaceutically active bindingpolypeptide after 180 minutes from start of release.

More suitably, when assayed in the Dynamic Dissolution Test, thepharmaceutical composition releases:

(i) 12-25% by weight of the pharmaceutically active binding polypeptideafter 60 minutes from start of release,

(ii) 45-58% by weight of the pharmaceutically active binding polypeptideafter 120 minutes from start of release and

(iii) 65% by weight or greater of the pharmaceutically active bindingpolypeptide after 180 minutes from start of release.

On occasion, a small premature release of polypeptide can occurimmediately on addition to Hanks buffer, followed by substantial releasecommencing shortly after. “Start of substantial release” can thereforebe defined as the point at which greater than 1% of the pharmaceuticallyactive binding polypeptide has been released and wherein a progressiveincrease in the amount of released pharmaceutically active bindingpolypeptide occurs at each of the time points 10, 20 and 30 minutesthereafter.

Suitably, when assayed in the Dynamic Dissolution Test, thepharmaceutical composition releases:

(i) 10-30% by weight of the pharmaceutically active binding polypeptideafter 60 minutes from start of substantial release,

(ii) 40-70% by weight of the pharmaceutically active binding polypeptideafter 120 minutes from start of substantial release and

(iii) 60% by weight or greater of the pharmaceutically active bindingpolypeptide after 180 minutes from start of substantial release.

Measurement of the Quantity of Released Pharmaceutically Active BindingPolypeptide (% Dissolution)

Measurement of % dissolution of pharmaceutically active bindingpolypeptide in the tests above may for example be performed by UV orHPLC analysis of the dissolution medium. Suitably, in the testsdescribed above, measurement of % dissolution of pharmaceutically activebinding polypeptide is performed by UV analysis of the dissolutionmedium. Suitably a UV path length of 10 mm and a spectrometer wavelengthof 279 nm is used. The absorbance of standard solutions containing knownquantities of polypeptide are measured to produce a standard curve, fromwhich the % of released pharmaceutically active binding polypeptide isthen ascertained. Alternatively, it may be assumed that this curve islinear and a single point determination of the standard may be made.

Without being bound by theory, it is believed that a gelling phenomenoncaused on contact of the pharmaceutical composition with an aqueousenvironment contributes to the sustained release profile of thepharmaceutically active binding polypeptide incorporated in thecomposition of the invention, such that the polypeptide is released intothe dissolution medium or in vivo environment at a slower and moreconsistent rate than would be expected from a compound with high watersolubility.

Intestinal Transit Times

Approximate human intestinal transit times (hours) in the fasting statethrough each region of the small intestine are as follows:

Through duodenum 0.3 Through jejunum 1.7 Through ileum 1.3

Transit times in the fed state are similar to those above. In light ofthese transit times, a composition with a pH sensitive enteric coatingformulated such that polypeptide starts to be released afterapproximately 18 minutes from entering the higher pH buffer environmentin vitro may be expected to first release polypeptide in vivo in thejejunum. Similarly, a composition with a pH sensitive enteric coatingformulated such that polypeptide starts to be released afterapproximately 120 minutes from entering the higher pH buffer environmentin vitro may be expected to first release polypeptide in vivo in theileum. Release would continue and then plateau while the compositionpasses through the remaining lower regions of the intestinal tract.

Tablet Components

Pharmaceutically Active Binding Polypeptides

Polypeptides are organic polymers consisting of a number of amino acidresidues bonded together in a chain. As used herein, ‘polypeptide’ isused interchangeably with ‘protein’ and ‘peptide’. Polypeptides are saidto be binding polypeptides when they contain one or more stretches ofamino acid residues which form an antigen-binding site, capable ofbinding to an epitope on a target antigen with an affinity (suitablyexpressed as a Kd value, a Ka value, a kon-rate and/or a koff-rate, asfurther described herein). ‘Binding polypeptide’ and ‘antigen-bindingpolypeptide’ are used synonymously herein. A binding polypeptide ispharmaceutically active if the binding polypeptide is capable ofexerting a beneficial pharmacological effect upon administration to asubject. Suitably a polypeptide is a pharmaceutically active bindingpolypeptide such that the polypeptide binds to, and more suitablyantagonises or neutralises, a biological target (typically a proteinsuch as a receptor, ion channel, enzyme, structural protein orcytokine). In some embodiments the pharmaceutically active bindingpolypeptide may agonise the biological target (such as a receptor).Pharmaceutically active binding polypeptides may include polypeptidessuch as antibodies (which are further described below), antibodiesmodified to comprise additional binding regions, antibody mimetics andantigen-binding antibody fragments (which are further described below).Further pharmaceutically active binding polypeptides may include DARPins(Binz et al. Journal of Molecular Biology 332(2):489-503), Affimers™,Fynomers™, Centyrins, Nanofitins® and cyclic peptides.

A conventional antibody or immunoglobulin (Ig) is a protein comprisingfour polypeptide chains: two heavy (H) chains and two light (L) chains.Each chain is divided into a constant region and a variable domain. Theheavy chain variable domains are abbreviated herein as VHC, and thelight (L) chain variable domains are abbreviated herein as VLC. Thesedomains, domains related thereto and domains derived therefrom, arereferred to herein as immunoglobulin chain variable domains (“ICVDs”).The VHC and VLC domains can be further subdivided into regions ofhypervariability, termed “complementarity determining regions” (“CDRs”),interspersed with regions that are more conserved, termed “frameworkregions” (“FRs”). The framework and complementarity determining regionshave been precisely defined (Kabat et al., 1991, herein incorporated byreference in its entirety). In a conventional antibody, each VHC and VLCis composed of three CDRs and four FRs, arranged from amino-terminus tocarboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4. The conventional antibody tetramer of two heavyimmunoglobulin chains and two light immunoglobulin chains is formed withthe heavy and the light immunoglobulin chains inter-connected by e.g.disulfide bonds, and the heavy chains similarity connected. The heavychain constant region includes three domains, CH1, CH2 and CH3. Thelight chain constant region is comprised of one domain, CL. The variabledomain of the heavy chains and the variable domain of the light chainsare binding domains that interact with an antigen. The constant regionsof the antibodies typically mediate the binding of the antibody to hosttissues or factors, including various cells of the immune system (e.g.effector cells) and the first component (C1q) of the classicalcomplement system. The term antibody includes immunoglobulins of typesIgA, IgG, IgE, IgD, IgM (as well as subtypes thereof), wherein the lightchains of the immunoglobulin may be kappa or lambda types. The overallstructure of immunoglobulin-gamma (IgG) antibodies assembled from twoidentical heavy (H)-chain and two identical light (L)-chain polypeptidesis well established and highly conserved in mammals (Padlan 1994).

An exception to conventional antibody structure is found in sera ofCamelidae. In addition to conventional antibodies, these sera possessspecial IgG antibodies. These IgG antibodies, known as heavy-chainantibodies (HCAbs), are devoid of the L chain polypeptide and lack thefirst constant domain (CH1). At its N-terminal region, the H chain ofthe homodimeric protein contains a dedicated immunoglobulin chainvariable domain, referred to as the VHH, which serves to associate withits cognate antigen (Muyldermans 2013, Hamers-Casterman et al., 1993,Muyldermans et al., 1994, herein incorporated by reference in theirentirety).

An antigen-binding antibody fragment (or “antibody fragment”,“immunoglobulin fragment” or “antigen-binding fragment”) as used hereinrefers to a portion of an antibody that specifically binds to a target(e.g. a molecule in which one or more immunoglobulin chains is not fulllength, but which specifically binds to a target). Examples of fragmentsencompassed within the term antigen-binding antibody fragment include:

(i) a Fab fragment (a monovalent fragment consisting of the VLC, VHC, CLand CH1 domains);

(ii) a F(ab′)2 fragment (a bivalent fragment comprising two Fabfragments linked by a disulfide bridge at the hinge region);

(iii) a Fd fragment (consisting of the VHC and CH1 domains);

(iv) a Fv fragment (consisting of the VLC and VHC domains of a singlearm of an antibody);

(v) an scFv fragment (consisting of VLC and VHC domains joined, usingrecombinant methods, by a synthetic linker that enables them to be madeas a single protein chain in which the VLC and VHC regions pair to formmonovalent molecules);

(vi) a VH (an immunoglobulin chain variable domain consisting of a VHCdomain (Ward et al., 1989));

(vii) a VL (an immunoglobulin chain variable domain consisting of a VLCdomain);

(viii) a V-NAR (an immunoglobulin chain variable domain consisting of aVHC domain from chondrichthyes IgNAR (Roux et al., 1998 and Griffiths etal., 2013, herein incorporated by reference in their entirety))

(ix) a VHH.

The total number of amino acid residues in a pharmaceutically activebinding polypeptide may be in the region of 50-3000, more suitably100-1500, more suitably 100-1000, more suitably 100-500, more suitably100-200. The total number of amino acid residues in a VHH or VH may bein the region of 110-140, is suitably 112-130, and more suitably115-125.

The examples provided herein relate to compositions comprisingimmunoglobulin chain variable domains per se. The principles of theinvention disclosed herein are, however, equally applicable to acomposition according to the invention comprising any pharmaceuticallyactive binding polypeptide. For example, the anti-TNF-alphaimmunoglobulin chain variable domains featured in the examples disclosedherein may be incorporated into a polypeptide such as a full lengthantibody. Such an approach is demonstrated by McCoy et al., 2014, whoprovide an anti-HIV VHH engineered as a fusion with a human Fc region(including hinge, CH2 and CH3 domains), expressed as a dimer construct.

Suitably, the pharmaceutically active binding polypeptide consists of animmunoglobulin chain variable domain. Suitably, the pharmaceuticallyactive binding polypeptide is an antibody or an antibody fragment.Suitably the antibody fragment is a VHH, a VH, a VL, a V-NAR, a Fabfragment, a VL or a F(ab′)2 fragment (such as a VHH or VH, most suitablya VHH).

Suitably the molecular weight of the pharmaceutically active bindingpolypeptide is 1-200 kD. More suitably 5-200 kDa, more suitably 10-200kDa, more suitably 10-180 kDa, more suitably 10-150 kDa, more suitably10-100 kDa, more suitably 10-50 kDa, more suitably 10-20 kDa, moresuitably 12-15 kDa, more suitably about 13 kDa.

Suitably the pharmaceutically active binding polypeptide has an aqueoussolubility of greater than 1 mg/mL, more suitably greater than 5 mg/mL,more suitably greater than 10 mg/mL, more suitably greater than 15mg/mL, more suitably greater than 20 mg/mL, more suitably greater than25 mg/mL, more suitably greater than 30 mg/mL.

Suitably the isoelectric point of the pharmaceutically active bindingpolypeptide is 6-8, more suitably 6.5-7.5, more suitably 6.7-6.9, moresuitably about 6.8.

SEQ ID Nos: 1 to 28 are polypeptide sequences of specific ICVDs whichare exemplary pharmaceutically active binding polypeptides according tothe invention. Suitably the pharmaceutically active binding polypeptidecomprises or more suitably consists of any one or more of SEQ ID Nos: 1to 28.

Suitably the pharmaceutically active binding polypeptide is for localdelivery to a region of the intestinal tract. Suitably thepharmaceutically active binding polypeptide is for local therapeuticeffect. “Local therapeutic effect” is defined as a beneficial biologicalimpact which only takes place, or is only appreciable, in the region towhich a pharmaceutically active binding polypeptide was first delivered.Local therapeutic effect excludes systemic effects (beneficial orotherwise). For example, a pharmaceutically active binding polypeptidewhich targets TNF-alpha, when delivered in a composition of theinvention, suitably binds to and neutralises the effects of TNF-alphapresent in the region(s) of the intestinal tract in which it was firstreleased. This pharmaceutically active binding polypeptide does not,however, significantly bind to significant quantities of TNF-alpha inother regions of the body and therefore does not have a significantsystemic impact.

The pharmaceutical composition of the invention provides protection to apharmaceutically active binding polypeptide during transit through thestomach and suitably also the duodenum, until pharmaceutically activebinding polypeptide is released in target regions of the intestinaltract. Accordingly, particularly suitable pharmaceutically activebinding polypeptides are those which are substantially inactivated whenexposed to the stomach and/or duodenum and which are therefore protectedfrom inactivation by the pH sensitive enteric coating of the compositionof the invention.

It is possible that a crude sample of polypeptide may contain impurities(such as inactive polypeptide) such that only a proportion of apolypeptide sample will be pharmaceutically active binding polypeptide.

The pharmaceutically active binding polypeptide can be in the form of apharmaceutically acceptable salt. Suitably the pharmaceutically activebinding polypeptide is not insulin. Suitably the pharmaceutically activebinding polypeptide is intra-granular.

Pharmaceutically active binding polypeptides can be obtained andmanipulated using the techniques disclosed for example in Green andSambrook 2012. For example, immunoglobulin chain variable domains may beobtained by preparing a nucleic acid encoding an immunoglobulin chainvariable domain using techniques for nucleic acid synthesis, followed byexpression of the nucleic acid thus obtained.

Specificity and Affinity

Specificity refers to the number of different types of antigens orantigenic determinants to which a particular antigen-binding polypeptidecan bind. The specificity of an antigen-binding polypeptide is theability of the antigen-binding polypeptide to recognise a particularantigen as a unique molecular entity and distinguish it from another.

Affinity, represented by the equilibrium constant for the dissociationof an antigen with an antigen-binding polypeptide (Kd), is a measure ofthe binding strength between an antigenic determinant and anantigen-binding site on the antigen-binding polypeptide: the lesser thevalue of the Kd, the stronger the binding strength between an antigenicdeterminant and the antigen-binding polypeptide (alternatively, theaffinity can also be expressed as the affinity constant (Ka), which is1/Kd). Affinity can be determined by known methods, depending on thespecific antigen of interest.

Suitably, a pharmaceutically active binding polypeptide will bind with adissociation constant (Kd) of at least 1×10⁻⁶ M, more suitably at least1×10⁻⁷ M, more suitably at least 1×10⁻⁸ M, more suitably at least 1×10⁻⁹M.

Any Kd value less than 10⁻⁶ is considered to indicate binding. Specificbinding of pharmaceutically active binding polypeptide to an antigen orantigenic determinant can be determined in any suitable known manner,including, for example, Scatchard analysis and/or competitive bindingassays, such as radioimmunoassays (RIA), enzyme immunoassays (EIA) andsandwich competition assays, and the different variants thereof known inthe art.

An anti-target polypeptide, a polypeptide which interacts with a target,or a polypeptide against a target, are all effectively polypeptideswhich bind to a target. A polypeptide may bind to a linear orconformational epitope.

Suitably the pharmaceutically active antigen binding polypeptide bindsto a target in the intestinal tract, such as a target in one or moreregions of the intestinal tract, such as an interleukin (such as IL-1,IL-1ra, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13,IL-15, IL-17, IL-18 and IL-23), an interleukin receptor (such as IL-6Rand IL-7R), a transcription factor (such as NF-kB), a cytokine (such asTNF-alpha, IFN-gamma TGF-beta and TSLP), a transmembrane protein (suchas gp130 and CD3), a surface glycoprotein (such as CD4, CD20, CD40), asoluble protein (such as CD40L), an integrin (such as a4b7 andAlphaEbeta7), an adhesion molecule (such as MAdCAM), a chemokine (suchas IP10 and CCL20), a chemokine receptor (such as CCR2 and CCR9), aninhibitory protein (such as SMAD7), a kinase (such as JAK3), a Gprotein-coupled receptor (such as sphingosine-1-P receptor) and productsof gastrointestinal pathogens.

Linkers and Multimers

A pharmaceutically active binding polypeptide may be a constructcomprising multiple polypeptides and therefore may suitably bemultivalent. Such a construct may comprise at least two identicalpolypeptides. Alternatively, a construct may comprise at least twopolypeptides which are different. Constructs can be multivalent and/ormultispecific. A multivalent construct (such as a bivalent construct)comprises two or more binding polypeptides and therefore presents two ormore sites at which attachment to one or more antigens can occur. Amultispecific construct (such as a bispecific construct) comprises twoor more different binding polypeptides which present two or more sitesat which either (a) attachment to two or more different antigens canoccur or (b) attachment to two or more different epitopes on the sameantigen can occur. A multispecific construct is multivalent.

The polypeptides in the construct can be linked to each other directly(i.e. without use of a linker) or via a linker. Suitably, the linker isa protease-labile or a non-protease-labile linker.

The linker is suitably a polypeptide and will be selected so as to allowbinding of the polypeptides to their epitopes. If used for therapeuticpurposes, the linker is suitably non-immunogenic in the subject to whichthe polypeptides are administered. Suitably the polypeptides are allconnected by non-protease-labile linkers. Suitably thenon-protease-labile linkers are of the format (G₄S)_(x). Suitably x is 1to 10, most suitably x is 6. Suitably the protease-labile linker is ofthe format [-(G_(a)S)_(x)—B-(G_(b)S)_(y)—]_(z) wherein a is 1 to 10; bis 1 to 10; x is 1 to 10; y is 1 to 10, z is 1 to 10 and B is K or R.Suitably a is 2 to 5, more suitably a is 4. Suitably b is 2 to 5, moresuitably b is 4. Suitably x is 1 to 5, more suitably x is 1. Suitably yis 1 to 5, more suitably y is 1. Suitably z is 1 to 3, more suitably zis 1. Suitably B is K. Capital lettering in the passage above refers tothe single letter amino acid code.

pH Sensitive Enteric Coatings

The pharmaceutical composition of the invention is provided with a pHsensitive enteric coating. Materials used for enteric coatings includefatty acids, waxes, shellac, plastics, and plant fibers. Suitably the pHsensitive enteric coating releases pharmaceutically active bindingpolypeptide when exposed to a region of the intestinal tract. Suitablythe region of the intestinal tract is a region of the small and/or largeintestine such as a region selected from at least one of the duodenum,jejunum, ileum, cecum, colon, rectum and anal canal. More suitably theregion is selected from at least one of the duodenum, jejunum and ileum.

The thickness of the pH sensitive enteric coating is selected such thatthe coating remains intact for the desired period of time during transitthrough the gastrointestinal tract and in particular for the desiredperiod of time following exposure to a pH which results in dissolutionof the coat. Suitably the thickness of the pH sensitive enteric coating(e.g. a pH sensitive enteric coating comprising poly(methacrylicacid-co-methyl methacrylate) 1:1 is 10-300 um, such as 50-200 um, suchas 70-170 um, such as 100-170 um.

Suitably, the quantity of pH sensitive enteric coating used in acomposition of the invention is present at 10-30%, more suitably 15-26%,more suitably 17-23%, more suitably 18-22%, more suitably about 20% w/wof the composition as a whole. Suitably these quantities are in thecontext of a 3 mm diameter compressed core.

Suitably the quantity of pH sensitive enteric coating used in acomposition of the invention is present at 15-35%, more suitably 20-30%,more suitably 22-28%, more suitably 24-26%, more suitably about 25% w/wof the core. Suitably these quantities are in the context of a 3 mmdiameter compressed core.

Alternatively, the quantity of pH sensitive enteric coating used in acomposition of the invention may be defined by the % weight gained bythe compressed core (or suitably the sub-coated compressed core) uponaddition of a pH sensitive enteric coating. In the Examples it isdemonstrated that a weight gain of 25% w/w of pH sensitive entericcoating resulted in the coating coming off after approximately 2 hoursduring the Dynamic Dissolution Test and that a weight gain of 17% w/w ofpH sensitive enteric coating resulted in the coating coming off afterapproximately 90 minutes during the Dynamic Dissolution Test. Thereforea range of 14%-30%, or more suitably 17%-27%, or more suitably 20-27%weight gained upon coating with pH sensitive enteric coating shouldprovide an optimal coating for dissolution in the small intestine.Furthermore, more specifically and if desired, 17%-20% weight gainedupon coating with pH sensitive enteric coating should provide an optimalcoating for dissolution in the duodenum or 20%-27% weight gained uponcoating with pH sensitive enteric coating should provide an optimalcoating for dissolution in the ileum. Suitably these quantities are inthe context of a 3 mm diameter compressed core.

If a compressed core has a cylindrical profile (such as a cylindricalminitablet), thickness measurements are suitably obtained by slicing themini-tablets both axially and radially (ignoring the thickness of thecoating on the corners of the tablets).

pH Sensitive Enteric Polymer Coat

The pH sensitive enteric coating may comprise a pH sensitive entericpolymer coat. A pH sensitive enteric polymer coat is a polymer which isincluded in the pH sensitive enteric coat and which acts as a barrier toprotect the polypeptide from the low pH of the stomach and suitably alsothe duodenum. A pH sensitive enteric polymer coat is insoluble at thehighly acidic pH found in the stomach, but dissolves rapidly at a lessacidic pH. Thus, suitably, the pH sensitive enteric polymer coat willnot dissolve in the acidic juices of the stomach (pH 1.5-4), but will doso in the higher pH environment present in the small intestine (pH above6) or in the colon (pH above 7.0). The pH sensitive enteric polymer coatis selected such that the polypeptide will start to be released at aboutthe time that the dosage reaches the small intestine, particularly theduodenum, jejunum and ileum; most suitably when the dosage reaches thejejunum. Suitably the pH sensitive enteric polymer coat does notdissolve until after 2 hours or longer exposure (suitably at least 6hours, more suitably 2-4 hours, more suitably 2-3 hours) to a pH of0.5-3.5, more suitably 0.6-3.0, more suitably 0.7-2.5, more suitably0.8-2.0, more suitably 0.9-1.5, more suitably about or exactly 1. Theskilled person will appreciate that proton pump inhibitors, H2inhibitors and acid neutralisers may raise stomach pH to approximately 4or even greater and therefore a pH sensitive enteric polymer coat whichdissolves at a higher pH may appropriately be used in a composition ofthe invention if taken simultaneously with proton pump inhibitors.Similarly, the skilled person will appreciate that a subject sufferingfrom e.g. achlorhydria will have a raised stomach pH (of greater than 5)and therefore a pH sensitive enteric polymer coat which dissolves at ahigher pH may appropriately be used in a composition of the invention ifadministered to a subject suffering from such a condition.

Suitably the pH sensitive enteric polymer coat comprises one or more of:methyl acrylate-methacrylic acid copolymers, cellulose acetatesuccinate, hydroxy propyl methyl cellulose phthalate, hydroxy propylmethyl cellulose acetate succinate (hypromellose acetate succinate),polyvinyl acetate phthalate (PVAP), methyl methacrylate-methacrylic acidcopolymers, sodium alginate and stearic acid. More suitably the pHsensitive enteric polymer coat comprises or consists of an anioniccopolymer based on methacrylic acid and methyl methacrylate. Moresuitably the pH sensitive enteric polymer coat comprises or consists ofpoly(methacrylic acid-co-methyl methacrylate) 1:1 (e.g. EudragitL100/L12.5) or poly(methacrylic acid-co-ethyl acrylate) 1:1 (e.g.Eudragit L100-55/L30-D55); most suitably poly(methacrylic acid-co-methylmethacrylate) 1:1.

The quantity of pH sensitive enteric polymer coat used in a compositionof the invention may be expressed as a percentage w/w of the pHsensitive enteric coating (i.e. the total solids of the pH sensitiveenteric coating). Suitably the pH sensitive enteric polymer coat ispresent at 40-70%, more suitably 55-65% by weight relative to the pHsensitive enteric coating.

The quantity of pH sensitive enteric polymer coat used in a compositionof the invention may also be expressed as a percentage w/w of thecomposition as a whole. Suitably, the quantity of pH sensitive entericpolymer coat used in a composition of the invention is present at10-30%, more suitably about 12% w/w of the composition as a whole.

If an entirely aqueous coating method is used to produce the compositionof the invention as opposed to a mixed aqueous and organic solventmethod then, in order to achieve an equivalent release profile, thequantity of pH sensitive enteric polymer coating should be increasedrelative to that used in an aqueous and organic solvent method.

Further Possible Components of the pH Sensitive Enteric Coat

Suitably the pH sensitive enteric coating comprises or consists of a pHsensitive enteric polymer coat optionally together with one or more of aplasticiser, an anti-tacking agent and a surfactant. More suitably thepH sensitive enteric coating consists of a pH sensitive enteric polymercoat, a plasticiser, an anti-tacking agent and a surfactant.

Suitably the pH sensitive enteric coating comprises a plasticiserwherein the plasticiser is triethyl citrate, the anti-tacking agent istalc and/or the surfactant is sodium lauryl sulphate. Suitably theplasticiser is present at 5-20% by weight relative to the pH sensitiveenteric coating and/or the anti-tacking agent is present at 20-40% byweight relative to the pH sensitive enteric coating and/or thesurfactant is present at 0.05-0.5% by weight relative to the pHsensitive enteric coating.

Suitably the pH sensitive enteric coating comprises, essentiallyconsists of, or consists of the following: 50-70% one or more pHsensitive enteric polymer coat, 7-17% one or more plasticisers, 20-40%one or more anti-tacking agents and 0.05-0.2% surfactant; all by weightrelative to the weight of the pH sensitive enteric coating.

Suitably a composition of the invention may comprise a sub-coatingbetween the compressed core and the pH sensitive enteric coating. Such asub-coating may improve adherence of the pH sensitive enteric coating tothe core. Suitably the sub-coating comprises or consists ofhydroxypropylmethylcellulose.

Excipients

The pharmaceutical composition of the invention suitably comprises atleast one excipient. Suitably the at least one excipient is selectedfrom one or more of: (i) one or more compression aids, (ii) one or moredisintegrants, (iii) one or more lubricants, (iv) one or more glidants,(v) one or more diluents and (vi) one or more binders. These excipientsare detailed as follows. % values expressed below are expressed as %weight of the compressed core.

Compression Aids

Compression aids serve to bind the components of the core togethergiving form and mechanical strength. Suitably the one or morecompression aids are intra-granular and extra-granular.

Suitably the composition of the invention comprises one or morecompression aids. Suitably the one or more compression aids is selectedfrom the list consisting of synthetic polymers such as crospovidone,saccharides such as sucrose, glucose, lactose and fructose, sugaralcohols such as mannitol, xylitol, maltitol, erythritol, sorbitol,water-soluble polysaccharides such as celluloses such as crystallinecellulose, microcrystalline cellulose, powdered cellulose,hydroxypropylcellulose and methyl cellulose, starches, syntheticpolymers such as polyvinylpyrrolidone, sodium starch glycolate,crospovidone and inorganic compounds such as calcium carbonate.

Suitably the one or more compression aids is present at 20-55%, such as30-40% by weight relative to the weight of the core.

It has been found that, surprisingly, mannitol improves the thermalstability of the pharmaceutically active binding polypeptide detailed inExample 1 (see Example 6). Suitably therefore, the one or morecompression aids is mannitol.

Binders

Binders are similar in function to compression aids. Suitably the coreof the composition of the invention comprises one or more binders.Suitably the one or more binders are intra-granular and extra-granular.Suitably the one or more binders is selected from the list consisting ofacacia, alginic acid, ammonio methacrylate copolymer, ammoniomethacrylate copolymer dispersion, carbomer copolymer, carbomerhomopolymer, carbomer interpolymer, carboxymethylcellulose sodium,microcrystalline cellulose, copovidone, sucrose, dextrin,ethylcellulose, gelatin, glucose, guar gum, low-substitutedhydroxypropyl cellulose, hypromellose, hydromellose acetate succinate,maltodextrin, maltose, methylcellulose, polyethylene oxide, povidone,starch such as corn starch; potato starch; pregelatinized starch;tapioca starch; wheat starch.

Suitably the one or more binders are present at 20-55%, such as 30-40%by weight relative to the weight of the core. Suitably the one or morebinders and compression aids are present at 20-55%, such as 30-40% byweight relative to the weight of the core.

Disintegrants

Disintegrants serve to aid dispersion of the core in thegastrointestinal tract, assisting to release the pharmaceutically activebinding polypeptide and increasing the surface area for dissolution.Disintegrants include super disintegrants. Suitably the core of thecomposition of the invention comprises one or more disintegrants.Suitably the one or more disintegrants are extragranular.

It is possible for disintegrants to counteract the effect of polypeptidegelling and as such disintegrants may be used to finely modulate therelease profile of a composition of the invention.

Suitably the one or more disintegrants is selected from the listconsisting of carboxymethyl cellulose, sodium carboxymethyl cellulose,croscarmellose sodium, cellulose such as low substitution degreehydroxypropylcellulose, starch such as sodium carboxymethyl starch,hydroxypropyl starch, rice starch, wheat starch, potato starch, maizestarch, partly pregelatinized starch. Suitably the disintegrant iscroscarmellose sodium. Suitably the one or more disintegrants is presentat 2-6%, such as about 4% by weight relative to the weight of the core.

It has been found that, surprisingly, croscarmellose sodium improves thethermal stability of the pharmaceutically active binding polypeptidedetailed in Example 1 (see ‘ID’, Example 6). Suitably therefore, the oneor more disintegrants is croscarmellose sodium.

Glidants

Glidants improve the flow of powders during tablet manufacturing byreducing friction and adhesion between particles. The core of thecomposition of the invention may comprise one or more glidants. Ifpresent, the glidants may be intra-granular or extra-granular. Suitablythe one or more glidants, if present, is selected from the listconsisting of calcium silicate, magnesium silicate, colloidal silicondioxide and talc. Most suitably the one or more glidants, if present, iscolloidal silicon dioxide. Suitably the one or more glidants, ifpresent, is present at 0.1-1.0%, more suitably about 0.5% by weightrelative to the weight of the core.

Lubricants

Lubricants have a similar action to glidants. Lubricants are addedprimarily to prevent sticking of the punches in the die duringtabletting. They may also slow disintegration and dissolution. Suitablythe core of the composition of the invention comprises one or morelubricants. Suitably the lubricants are intra-granular andextra-granular. Suitably the one or more lubricants is selected from thelist consisting of glyceryl behenate, a stearic acid salt such ascalcium stearate; magnesium stearate; zinc stearate, mineral oil,polyethylene glycol, sodium lauryl sulfate, sodium stearyl fumarate,starch such as corn starch; potato starch; pregelatinized starch;tapioca starch; wheat starch, stearic acid, talc, vegetable oil and zincstearate.

Suitably the core comprises one or more lubricants wherein the one ormore lubricants is present at 0.1-2%, such as about 1% by weightrelative to the weight of the core.

It has been found that, surprisingly, magnesium stearate improves thethermal stability of the pharmaceutically active binding polypeptidedetailed in Example 1 (see ‘E’, Example 6). Suitably therefore, the oneor more lubricants is magnesium stearate.

Other Components

The advantageous release profiles of compositions of the invention canbe achieved without addition of substances which conventionally delaythe release of pharmaceutically active binding polypeptide, or protectpharmaceutically active binding polypeptide, such as hydrogels.Hydrogels are three-dimensional mesh like networks containinghydrophilic polymers that imbibe substantial quantities of water andform a gel like matrix as a result of physical or chemical cross linkingof individual polymer chains. Suitably the composition of the inventioncomprises less than 1%, more suitably less than 0.5%, more suitably lessthan 0.1% by weight hydrogels, more suitably the composition of theinvention comprises no hydrogels. Furthermore, the advantageous releaseprofiles of compositions of the invention may be achieved withoutincluding alginates, hydroxypropyl methylcellulose (HPMC) and/orcopolymers based on methacrylic acid and methyl methacrylate in thecompressed core of the composition. The advantageous release profiles ofcompositions of the invention are furthermore suitably achieved withoutaddition of sustained release coatings (e.g. a coat permitting diffusionof active agent, which may be positioned between the pH sensitiveenteric coat and the compressed core). Similarly, the compositions ofthe invention suitably do not comprise any one or more of the followingcomponents which can be added to formulations to achieve absorption ofthe active ingredient: penetration/permeability enhancers (includingcell penetrating peptides or membrane modifying substances); carriersystems (including nanoparticles, cyclodextrins, polymeric carriers orlipid based systems); mucous modifying, adhesive or penetratingexcipients; or sugar micro needles.

Presentation Forms and Structure

The pharmaceutical compositions of the invention may be presented in avariety of forms. These include solid oral dosage forms comprising pHsensitive enteric coatings and compressed cores such as tablets ormini-tablets.

Mini-tablets may be round, cylindrical tablets or disc-like in shape.Mini-tablets are suitably 1 to 5 mm in diameter, more suitably 1-4 mm indiameter, more suitably 1-3 mm in diameter, more suitably 2-3 mm indiameter, more suitably about 3 mm in diameter, more suitably about 2 mmin diameter. Mini-tablets are typically produced by compression. Theyprovide a smooth substrate for enteric coating using e.g. eitherconventional perforated coating pans or fluid-bed apparatus.Mini-tablets offer finished dosage form flexibility in that they can bedelivered within capsules or sachets.

Such mini-tablets may be presented in a capsule. Capsules may behard-shelled capsules, which are typically made using gelatin or HPMC(most suitably HPMC) and contain dry, powdered ingredients or miniaturepellets made by e.g. processes of extrusion or spheronisation. These aremade in two halves: a lower-diameter “body” that is filled and thensealed using a higher-diameter “cap”. Alternatively capsules may besoft-shelled capsules. Both of these classes of capsules are made fromaqueous solutions of gelling agents, such as animal protein (mainlygelatin) or plant polysaccharides or their derivatives (such ascarrageenans and modified forms of starch and cellulose).

The pharmaceutical composition of the invention comprises a compressedcore and a pH-sensitive enteric coating, wherein the compressed corecomprises a pharmaceutically active binding polypeptide.

“Compressed” as used herein refers to a substance which has undergonecompression, i.e. squeezing or pressing.

Suitably the compressed core essentially consists of, or consists of oneor more compression aids; one or more disintegrants; one or morelubricants a pharmaceutically active binding polypeptide.

Suitably the hardness of the compressed core is 20-110 N, more suitably40-100 N, most suitably 60-90 N.

Suitably the compressed core comprises, essentially consists of, orconsists of the following: 20-55% one or more compression aids (e.g.mannitol and microcrystalline cellulose); 2-6% one or more disintegrants(e.g. croscarmellose sodium); 0.1-2% one or more lubricants (e.g.magnesium stearate) and 40-80% pharmaceutically active bindingpolypeptide (e.g. an immunoglobulin chain variable domain); all byweight relative to the weight of the core. More suitably the compressedcore comprises, essentially consists of, or consists of the following:30-40% one or more compression aids (e.g. mannitol and microcrystallinecellulose); 3-5% one or more disintegrants (e.g. croscarmellose sodium);0.5-1.5% one or more lubricants (e.g. magnesium stearate) and 50-70%pharmaceutically active binding polypeptide (e.g. an immunoglobulinchain variable domain); all by weight relative to the weight of thecore.

Suitably, the pharmaceutically active binding polypeptide is present at30-80%, such as 40-75%, such as about 50%-60% by weight relative to theweight of the core.

Therapeutic Use and Delivery

The pharmaceutical composition of the invention is suitably foradministration to a human. A therapeutically effective amount of apharmaceutical composition of the invention is an amount which iseffective, upon single or multiple dose administration to a subject, intreating or preventing disease in a subject. A therapeutically effectiveamount may vary according to factors such as the disease state, age,sex, and weight of the individual, and the ability of the pharmaceuticalcomposition to elicit a desired response in the individual. Atherapeutically effective amount is also one in which any toxic ordetrimental effects of the pharmaceutical composition are outweighed bythe therapeutically beneficial effects.

A pharmaceutical composition of the invention is formulated for oraldelivery. A key problem with oral delivery is ensuring that sufficientpharmaceutically active binding polypeptide reaches the area of theintestinal tract where it is required. Factors which prevent apolypeptide reaching the area of the intestinal tract where it isrequired include the presence of proteases in digestive secretions whichmay degrade the polypeptide. Suitably, the polypeptide is substantiallystable in the presence of one or more of such proteases by virtue of theinherent properties of the polypeptide itself. Suitably, the polypeptideis substantially stable when exposed to a region of the intestinaltract. Alternatively, a large dose of polypeptide may be administered tocompensate for quantities of polypeptide becoming degraded in theintestinal tract.

It is known that proteins delivered directly to the ileum can beimmunogenic whereas the same proteins delivered orally and subjected tostomach and duodenal digestion, induce immunological tolerance (seeMichael 1989). The compositions of the invention are enterically coatedresulting in, on oral administration, pharmaceutically active bindingpolypeptide being first exposed to the subject's immune system whenreleased in a region of the intestinal tract such as in the ileum. Theskilled person may view such a delivery method as immunologicallyequivalent to direct delivery to the ileum. It is surprising and highlyadvantageous therefore that an immunologically tolerant response hasbeen observed in respect of the orally-administered compositions of theinvention.

A surfactant may also be added to the pharmaceutical composition toreduce aggregation of the polypeptide and/or minimize the formation ofparticulates in the formulation and/or reduce adsorption. Exemplarysurfactants include polyoxyethylensorbitan fatty acid esters (Tween),polyoxyethylene alkyl ethers (Brij), alkylphenylpolyoxyethylene ethers(Triton-X), polyoxyethylene-polyoxypropylene copolymer (Poloxamer,Pluronic), and sodium dodecyl sulfate (SDS). Examples of suitablepolyoxyethylenesorbitan-fatty acid esters are polysorbate 20, andpolysorbate 80. Exemplary concentrations of surfactant may range fromabout 0.001% to about 10% w/v.

The pharmaceutically active binding polypeptide may be lyophilised. Alyoprotectant may be added in order to protect the polypeptide againstdestabilizing conditions during the lyophilization process. For example,known lyoprotectants include sugars (including glucose, sucrose, mannoseand trehalose); polyols (including mannitol, sorbitol and glycerol); andamino acids (including alanine, glycine and glutamic acid).Lyoprotectants can be included in an amount of about 10 mM to 500 mM.

Alternatively, the pharmaceutically active binding polypeptide may bespray dried.

The dosage ranges for administration of the pharmaceutical compositionof the invention are those to produce the desired therapeutic effect.The dosage range required depends on the precise nature of thepharmaceutical composition, the age of the patient, the nature, extentor severity of the patient's condition, contraindications, if any, andthe judgement of the attending physician. Variations in these dosagelevels can be adjusted using standard empirical routines foroptimisation.

Suitable human dosages of the pharmaceutically active bindingpolypeptide are in the range of 10 mg-5000 mg, such as 50 mg-1500 mg,such as 100 mg-500 mg. Suitable dosages of the pharmaceutically activebinding polypeptide per kg of body weight are in the range of 1 mg-500mg, such as 5 mg-150 mg, such as 10 mg-50 mg. The human dose or the doseper kg bodyweight may be administered daily or more frequently, forexample 2, 3 or 4 times per day or less frequently for example everyother day or once per week, once per fortnight or once per month. Moresuitably the dose is administered 3 times per day.

In one aspect of the invention there is provided the use of thepharmaceutical composition in the manufacture of a medicament for thetreatment of autoimmune and/or inflammatory diseases of the intestinaltract. In a further aspect of the invention there is provided a methodof treating autoimmune and/or inflammatory diseases of the intestinaltract comprising administering to a person in need thereof atherapeutically effective amount of the pharmaceutical composition.

The word ‘treatment’ is intended to embrace prophylaxis as well astherapeutic treatment. Treatment of diseases also embraces treatment ofexacerbations thereof and also embraces treatment of patients inremission from disease symptoms to prevent relapse of disease symptoms.

Diseases of the Intestinal Tract

Suitably the pharmaceutical composition of the invention is for use inthe treatment or prevention of diseases of the intestinal tract.Diseases of the intestinal tract relate to diseases affecting the smallintestine and large intestine. The small and large intestines may beaffected by infectious, autoimmune and other types of diseases.

Autoimmune Diseases and/or Inflammatory Diseases of the Intestinal Tract(IT)

The chronic inflammatory bowel diseases (IBD) Crohn's disease andulcerative colitis, which afflict both children and adults, are examplesof autoimmune and inflammatory diseases of the IT (Hendrickson et al.,2002, herein incorporated by reference in its entirety). Ulcerativecolitis is defined as a condition where the inflammatory response andmorphologic changes remain confined to the colon. The rectum is involvedin 95% of patients. Inflammation is largely limited to the mucosa andconsists of continuous involvement of variable severity with ulceration,edema, and hemorrhage along the length of the colon (Hendrickson et al.,2002, herein incorporated by reference in its entirety). Ulcerativecolitis is usually manifested by the presence of blood and mucus mixedwith stool, along with lower abdominal cramping which is most severeduring the passage of bowel movements. Clinically, the presence ofdiarrhoea with blood and mucus differentiates ulcerative colitis fromirritable bowel syndrome, in which blood is absent.

Unlike ulcerative colitis, the presentation of Crohn's disease isusually subtle, which leads to a later diagnosis. Factors such as thelocation, extent, and severity of involvement determine the extent ofgastrointestinal symptoms. Patients who have ileocolonic involvementusually have postprandial abdominal pain, with tenderness in the rightlower quadrant and an occasional inflammatory mass. Symptoms associatedwith gastroduodenal Crohn's disease include early satiety, nausea,emesis, epigastric pain, or dysphagia. Perianal disease is common, alongwith anal tags, deep anal fissures, and fistulae (Hendrickson et al.,2002, herein incorporated by reference in its entirety).

Suitably the polypeptide, pharmaceutical composition or construct of theinvention is for use in the treatment of an autoimmune and/orinflammatory disease of the IT selected from the list consisting ofCrohn's disease, ulcerative colitis, irritable bowel disease, diabetestype II, glomerulonephritis, autoimmune hepatitis, Sjogren's syndrome,celiac disease and drug- or radiation-induced mucositis; more suitablyCrohn's disease, ulcerative colitis, irritable bowel disease, celiacdisease and drug- or radiation-induced mucositis; more suitably Crohn'sdisease, ulcerative colitis and irritable bowel disease; most suitablyCrohn's disease.

Combination Therapy

It is within the scope of the invention to use the pharmaceuticalcomposition of the invention in therapeutic methods for the treatment ofautoimmune diseases as an adjunct to, or in conjunction with, otherestablished therapies normally used in the treatment of autoimmunediseases.

For the treatment of IBD (such as Crohn's disease or ulcerativecolitis), possible combinations include combinations with, for example,one or more active agents selected from the list comprising:5-aminosalicylic acid, or a prodrug thereof (such as sulfasalazine,olsalazine or bisalazide); corticosteroids (e.g. prednisolone,methylprednisolone, or budesonide); immunosuppressants (e.g.cyclosporin, tacrolimus, methotrexate, azathioprine or6-mercaptopurine); anti-IL-6R antibodes (e.g. tocilizumab), anti-IL-6antibodies, anti-TNF-alpha antibodies (e.g., infliximab, adalimumab,certolizumab pegol or golimumab); anti-IL12/IL23 antibodies (e.g.,ustekinumab); anti-IL6R antibodies or small molecule IL12/IL23inhibitors (e.g., apilimod); Anti-alpha-4-beta-7 antibodies (e.g.,vedolizumab); MAdCAM-1 blockers (e.g., PF-00547659); antibodies againstthe cell adhesion molecule alpha-4-integrin (e.g., natalizumab);antibodies against the IL2 receptor alpha subunit (e.g., daclizumab orbasiliximab); JAK3 inhibitors (e.g., tofacitinib or R348); Sykinhibitors and prodrugs thereof (e.g., fostamatinib and R-406);Phosphodiesterase-4 inhibitors (e.g., tetomilast); HMPL-004; probiotics;Dersalazine; semapimod/CPSI-2364; and protein kinase C inhibitors (e.g.AEB-071). The most suitable combination agents are tocilizumab,infliximab, adalimumab, certolizumab pegol or golimumab.

Hence another aspect of the invention provides a pharmaceuticalcomposition of the invention in combination with one or more furtheractive agents, for example one or more active agents described above.

In a further aspect of the invention, the pharmaceutical composition isadministered sequentially, simultaneously or separately with at leastone active agent selected from the list above.

Similarly, another aspect of the invention provides a combinationproduct comprising:

(A) a pharmaceutical composition of the present invention; and

(B) one or more other active agents,

wherein each of components (A) and (B) is formulated in admixture with apharmaceutically-acceptable adjuvant, diluent or carrier. In this aspectof the invention, the combination product may be either a single(combination) formulation or a kit-of-parts. Thus, this aspect of theinvention encompasses a combination formulation including apharmaceutical composition of the present invention and anothertherapeutic agent, in admixture with a pharmaceutically acceptableadjuvant, diluent or carrier.

The invention also encompasses a kit-of-parts comprising components:

(i) a pharmaceutical composition of the present invention in admixturewith a pharmaceutically acceptable adjuvant, diluent or carrier; and

(ii) a formulation including one or more other active agents, inadmixture with a pharmaceutically-acceptable adjuvant, diluent orcarrier, which components (i) and (ii) are each provided in a form thatis suitable for administration in conjunction with the other.

Component (i) of the kit of parts is thus component (A) above inadmixture with a pharmaceutically acceptable adjuvant, diluent orcarrier. Similarly, component (ii) is component (B) above in admixturewith a pharmaceutically acceptable adjuvant, diluent or carrier. The oneor more other active agents (i.e. component (B) above) may be, forexample, any of the agents mentioned above in connection with thetreatment of autoimmune diseases such as IBD (e.g. Crohn's diseaseand/or ulcerative colitis). If component (B) is more than one furtheractive agent, these further active agents can be formulated with eachother or formulated with component (A) or they may be formulatedseparately. In one embodiment component (B) is one other therapeuticagent. In another embodiment component (B) is two other therapeuticagents. The combination product (either a combined preparation orkit-of-parts) of this aspect of the invention may be used in thetreatment or prevention of an autoimmune disease (e.g. the autoimmunediseases mentioned herein).

Suitably the pharmaceutical composition of the invention is for use as amedicament and more suitably for use in the treatment of an autoimmuneand/or inflammatory disease.

Preparative Methods

Dry granulation followed by compression or alternatively directcompression may be used to manufacture the compressed core. Powders thatcan be mixed well do not require granulation and can be compressed intotablets through direct compression. These methods are particularlyappropriate if the composition of the invention is to be delivered as atablet, such as a mini-tablet.

Granulation processes create granules by light compaction of the powderblend under low pressures. The compacts so-formed are broken up gentlyto produce granules (agglomerates). This process is often used when theproduct to be granulated is sensitive to moisture and heat. Drygranulation can be conducted on a tablet press using slugging tooling oron a roll press called a roller compactor. Pressures may be varied toattain proper densification and granule formation.

Components may be added to the pharmaceutical composition duringmanufacture such that the components become extragranular orintragranular. If the component is mixed with other components prior togranulation, which is further prior to tablet compression to form thecore, the component will be incorporated into the granules and will thusbe intragranular. If the component is mixed with formed dry granulesbefore tablet compression to form the core, the component will beextragranular.

Compressed cores may be fabricated by direct compression by punches anddies fitted to a tabletting press, ejection or compression molding,granulation followed by compression, or forming a paste and extrudingthe paste into a mold or cutting the extrudate into short lengths andoptionally using a spheroniser to round the edges of the lengths soformed. Suitably, the process used for preparing compressed cores is drygranulation of the component mixture followed by compression, preferablyinto mini-tablets. Suitably compression is carried out using atabletting press such as a single punch machine (e.g. a Manesty F3) or arotary tablet press (e.g. a Manesty Technipress, suitably using a topcam compression force setting of 29).

In producing the pharmaceutical composition of the invention, suitablythe pharmaceutically active binding polypeptide is dry granulated toform granules. More suitably the pharmaceutically active bindingpolypeptide (e.g. an immunoglobulin chain variable domain) and one ormore compression aids (e.g. mannitol) are granulated together. Moresuitably the pharmaceutically active binding polypeptide (e.g. animmunoglobulin chain variable domain), one or more compression aids(e.g. mannitol) and one or more lubricants (e.g. magnesium stearate) aregranulated together. These components therefore become intra-granular.The granules so produced may then suitably be compressed to form thecompressed core.

Alternatively, after granulation, suitably the one or more compressionaids (e.g. microcrystalline cellulose), the one or more disintegrants(e.g. croscarmellose sodium) and the one or more lubricants (e.g.magnesium stearate) are blended with the granules, followed bycompression to form the compressed core.

As used herein, ‘infra-granular’ means present within granules (beforecompression) and ‘extra-granular’ means present between granules (aftercompression).

In one aspect of the invention there is provided a method of producingthe pharmaceutical composition of the invention wherein (i) apharmaceutically active binding polypeptide, one or more compressionaids and one or more lubricants are dry granulated together to formgranules, wherein the pharmaceutically active binding polypeptide is animmunoglobulin chain variable domain, the one or more compression aidsis mannitol and the one or more lubricants is magnesium stearate,followed by (ii) blending with the granules one or more compressionaids, one or more disintegrants and one or more lubricants, wherein theone or more compression aids is microcrystalline cellulose, the one ormore disintegrants is croscarmellose sodium and the one or morelubricants is magnesium stearate, followed by (iii) compressing theblend to form a compressed core; wherein the compressed core consists of20-55% mannitol and microcrystalline cellulose; 2-6% croscarmellosesodium; 0.1-2% magnesium stearate and 40-75% immunoglobulin chainvariable domain all by weight relative to the weight of the core,followed by (iv) coating the compressed core with a pH sensitive entericcoating. In a further aspect the invention provides a pharmaceuticalcomposition obtainable by the preceding method.

pH sensitive enteric coatings may be applied using organic solvent,using aqueous solution or using a mixture of organic solvent and water.Greater quantities of pH sensitive enteric polymer coat should beincluded when using an entirely aqueous solution coating process.

According to one aspect of the invention there is provided a method ofmaking the pharmaceutical composition according to the inventioncomprising compressing a blend of the pharmaceutically active bindingpolypeptide and the one or more excipients to form a core, followed bycoating the core with a pH sensitive enteric coating. Suitably the coreis coated in a sub-coat before coating with a pH sensitive entericcoating.

The present invention will now be further described by means of thefollowing non-limiting examples.

EXAMPLES Example 1: Formulation

A solid pharmaceutical composition according to the invention wasproduced in the form of mini-tablets by dry granulation and compression.The mini-tablets were then presented in different presentations, whereineach presentation contained a different quantity of mini tablets indifferent sizes of capsules. The main presentation used in the examplesdetailed below was a size 00 HPMC capsule containing 15 entericallycoated mini-tablets (total 185 mg of pharmaceutically active bindingpolypeptide). The mini-tablet cores had a diameter of 3 mm (excludingcoating thickness) and a hardness of approximately 90 N.

The components contained in each mini-tablet and therefore in the 15mini-tablets contained in the capsule are listed in Table 2 below.

TABLE 2 Quantity Quantity % (mg/capsule) (mg) w/w in 185 mg dose 12 mgdose Name of mini compo- (15 mini- (1 mini- tablet component Functionsition tablets) tablet) Mini-tablet cores Total polypeptide Active 45.7225 15 pharmaceu- tical ingre- dient (API) Mannitol Compression 12.059.25 3.95 aid Microcrystalline Compression 14.6 72 4.8 cellulose aidCroscarmellose Super 3.1 15 1 sodium disintegrant Magnesium stearateLubricant 0.8 3.75 0.25 Sub coating Hydroxypropyl- Polymer coat 3.818.75 1.25 methyl cellulose pH sensitive enteric coating Eudragit ® L100Enteric 11.7 57.76 3.85 polymer coat Triethyl citrate Plasticiser 2.311.51 0.77 Talc Anti-tacking 5.9 28.93 1.93 agent Sodium laurylSurfactant 0.04 0.20 0.01 sulphate

The total polypeptide in the composition has a purity of approximately70-90% such that 225 mg of polypeptide contains 185 mg ofpharmaceutically active binding polypeptide.

The pharmaceutically active binding polypeptide used throughout theexamples is one of the ICVDs recited in SEQ ID Nos: 1-28. This is a 115amino acid, 12.6 kDa polypeptide. The pI of the polypeptide is 6.8 andthe polypeptide has an aqueous solubility of greater than 30 mg/mL. TheICVD binds with high affinity to, and has potent neutralising activityagainst, human and Cynomolgus monkey TNF-alpha.

The mini-tablets were produced by the following methodology.

The lyophilised polypeptide was blended with mannitol and a portion ofthe magnesium stearate and dry slugged to increase its density. Thismaterial was then passed through a screen, blended with the othermini-tablet excipients (microcrystalline cellulose, croscarmellosesodium and the remaining magnesium stearate) and compressed to producethe mini-tablets. The mini-tablets were then coated with a 5% solutionof hydroxylpropyl methyl cellulose in ethanol:water 80:20, dried and thesolvent removed to create a sub-coat and a smoother surface. Themini-tablets were then coated with Eudragit® L100 polymer, together withtriethyl citrate, talc and sodium lauryl sulphate, as an organicsolution in isopropyl alcohol and water and dried to create apH-sensitive enteric coat, such that each mini-tablet gained 25% weight.The resulting approximately 3 mm diameter mini-tablets were then filledinto capsules the doses given above.

Two separate batches of mini-tablets were produced on differentoccasions. These batches are referred to herein as Batch A and Batch B.Both batches contain identical quantities of components as listed inTable 1. The pH sensitive enteric coating on Batch A had a thickness of100-170 um while the pH sensitive enteric coating on Batch B had athickness of 70-170 um.

Example 2: The Pharmacopeial Enteric Coating Test

Prior studies (not shown) established that the sub-coated compressedcore needed to gain greater than 17% additional weight on addition of pHsensitive enteric coating optimal release timing.

Six capsules of Example 1 containing mini-tablets from Batch A weresubjected to the Pharmacopeial Enteric Coating Test, one capsule pervessel.

Reference standards were prepared containing 0.0206 mg/mL and 0.206mg/mL of ICVD.

The capsules promptly dissolved as expected, releasing the mini-tabletsto the acid environment.

It was found that the mini-tablets released less than 10% of the ICVDduring the two hour period of the test (data not shown). The compositionis therefore compliant with the dissolution requirements fordelayed-release solid dosage forms administered orally set out inEuropean Pharmacopoeia 8.0, 2.9.3, “Dissolution test for solid dosageforms”.

During repeat testing, when the composition was left in the testingmedium with continued stirring for greater than 2 hours, it wasestablished by visual inspection (and by there being no increase inUV280 nm) that the enteric coat maintained integrity for greater than orequal to 6 hours.

Example 3: The Pharmacopeial Dissolution Test

After testing the mini-tablets from each of the six vessels in thePharmacopeial Enteric Coating Test as detailed in Example 2, themini-tablets from each of the six vessels were then tested using thePharmacopeial Dissolution Test, as detailed above. The results of thetest are shown in Table 3, FIG. 1 (data points for individual vessels1-6) and FIG. 2 (average for vessels 1-6, with the dissolution rangeswhich are recited under “Measurement of sustained release: ThePharmacopeial Dissolution Test” above, illustrated by arrows).

TABLE 3 Time (minutes) % Dissolution 5 0.6 10 1.5 15 2.3 30 16.6 45 29.060 41.6 90 63.1 120 71.3 150 73.5

In summary, it can be seen that sustained release of pharmaceuticallyactive binding polypeptide was achieved. Sustained release was achievedover the course of approximately 2 to 2.5 hours. This release profilewas expected to translate to an ideal sustained release profile in vivo.This expectation was confirmed by the in vivo experiments detailedbelow.

Example 4: The Dynamic Dissolution Test

The solid pharmaceutical composition detailed above under Example 1 wastested using the Dynamic Dissolution Test. Six samples, each samplecontaining 15 mini-tablets from Batch A and six samples, each samplecontaining 11 mini-tablets from Batch B, were tested. The conditionsused for each sample of 15 Batch A mini-tablets were in line with TheDynamic Dissolution Test detailed above. The conditions used for eachsample of 11 Batch B mini-tablets varied from The Dynamic DissolutionTest in that 800 mL of 0.1M HCl was used per sample.

No dissolution of ICVD from the mini-tablets took place during the acidstage for any sample of either Batch A (FIG. 4, curve preceding 2 hourpoint) or Batch B (FIG. 5, curve preceding 2 hour point).

These same mini-tablets were then transferred to Hanks buffer accordingto The Dynamic Dissolution Test. Transfer to Hanks buffer (2 hour point)is denoted by an upright dotted line in FIGS. 3, 4 and 5.

A stock and working solution was prepared. The stock solution wasprepared in the dissolution vessel filed with 798.5 mL of preconditioned Hanks hydrogen carbonate buffer solution of pH 5.79 byadding of 1.50 mL ICVD standard solution (known quantity andconcentration of ICVD). The samples of dissolution media were filtratedduring sampling over a 1 μm Poroplast-filter (PES, DissolutionAccessories, Amsterdam, Netherlands) and measured without furtherpre-treating. Amount of the dissolved drug was determined by means ofUV-Vis spectroscopy (Agilent 8453, Agilent Technologies, Santa Clara,USA) in close loop mode. The absorbance was measured using quartz flowthrough cells (Hellma, Müllheim, Germany) with 10 mm light path indifferential mode at 279 (signal) and 450-550 nm (background subtractionover range).

The amount of the dissolved ICVD was calculated individually based onthe mean standard absorbance according to the equation given below:ICVD dissolved=A*V*FWherein:A—Measured absorbance of the sampleV—Corrected volumeF—Calibration factor obtained based on the absorbance of the standardsolution

The volume correction of the sample and standard solution were performedindividually. The volume of the dissolution medium of the standard andtest solutions was measured at the end of the test. It was assumed thatthe volume contraction due to the evaporation was linear over time andthe volume of the sample and standard solutions were calculated for eachsampling point individually and were accordingly considered in thecalculations.

When testing the Batch A mini-tablet samples, it was found that greaterthan 1% of the polypeptide first started to be released after 1.67 hoursfrom first transfer to Hanks buffer (see FIG. 4, region after transferto Hanks buffer at 2 hours). When testing Batch B, it was found thatgreater than 1% of the polypeptide first started to be released after3.17 hours from first transfer to Hanks buffer (see FIG. 5, region aftertransfer to Hanks buffer at 2 hours).

This time point, at which greater than 1% of the pharmaceutically activebinding polypeptide has first started to be released, is the ‘start ofrelease’. The start of release is denoted by an arrow in FIGS. 4 and 5.These time periods for both batches are suitable for a delayed releaseproduct.

The % dissolution of ICVD following start of release was then recorded.The results for Batch A are shown in FIG. 4 (region following start ofrelease at 1.67 hours following first transfer to Hanks buffer at 2hours) and the results for Batch B are shown in FIG. 5 (region followingstart of release at 3.17 hours following first transfer to Hanks bufferat 2 hours).

A summary of the % dissolution of ICVD at time points 60, 120 and 180minutes after start of release is given in Table 4 below.

TABLE 4 Mean Mean results for results for % of ICVD released after startof release Batch A Batch B % released 60 mins after start of release14.3 23.1 % released 120 mins after start of release 47.1 57.7 %released 180 mins after start of release 67.6 68.4

After exiting the stomach, transit through the duodenum and jejunumtakes approximately 2 hours (see above under “Intestinal TransitTimes”). Accordingly, it was expected that the in vitro release profilesabove would translate to sustained release profiles in vivo whereinrelease starts to take place around the distal jejunum, withsubstantially complete release achieved in the colon. All released ICVDwould then be expected to travel through the remaining lower regions ofthe intestinal tract. This expectation was confirmed by the in vivoexamples below.

Example 5: Excipient Compatibility

The effect of the excipients used in the composition on the meltingtemperature (Tm) of the ICVD was investigated.

Thermal shift assays (TSA) are a common method used in proteinbiochemistry to examine the effect of solutes on a protein's structure.Certain solutes (salts, excipients etc) may interact with the proteinand cause either stabilisation or destabilisation of the protein. Thiseffect can be assessed by comparing the melting temperature (Tm) of theprotein with or without the compound in question. Increases in Tmindicate stabilisation, i.e., a strengthening of forces that hold theprotein in shape. Decreases in Tm indicate the reverse.

In this assay, the protein is mixed with the hydrophobic dye syproorange and heated gradually from 25° C. to 98° C. As the proteins melt(unfold, a.k.a. denature), their hydrophobic cores are exposed and syproorange binds to these residues via hydrophobic interactions. Syproorange fluoresces only when bound to the protein and, in this manner,the unfolding of the protein is measured in real time by lasers in theqPCR machine. The data from the machine is processed in Graphpad Prism,using Boltzmann curve fitting. The Tm is taken from the inflection pointof the Boltzmann curve. Excipients from were mixed in 1×PBS, pH 7.4, orwater by vortexing. Any insoluble material was removed by centrifugationand the supernatants taken for assay.

The following excipients were tested (labels refer to the legend in FIG.6).

A—(−0.1° C.) 8.88 uM ICVD; control in 1 (1×PBS pH 7.4)

B—(0.1° C.) 8.88 uM ICVD; 17 mg/ml Avicel PH102 in 1 (1×PBS pH 7.4)

C—(0.5° C.) 8.88 uM ICVD; 17 mg/ml Mannitol in 1 (1×PBS pH 7.4)

D—(0.6° C.) 8.88 uM ICVD; 8 mg/ml AcDiSol in 1 (1×PBS pH 7.4)

E—(1.2° C.) 8.88 uM ICVD; 2 mg/ml magnesium stearate in 1 (1×PBS pH 7.4)

F—(0.2° C.) 8.88 uM ICVD; 17 mg/ml Avicel PH112 in 1 (1×PBS pH 7.4)

Avicel PH102 and Avicel PH112 are types of microcrystalline celluloseand AcDiSol is a type of croscarmellose sodium.

The results are shown in FIG. 6. In summary, it was found that theseexcipients had no impact (detrimental or otherwise) on the Tm of theprotein, as most Tm curves produced with excipient exposure remainedsubstantially the same as control (control is shown as Tm curve ‘A’).Surprisingly, it was noted that small improvements in thermal stabilityof the protein were achieved on exposure to mannitol (+0.5° C.),AciDiSol (+0.6° C.) and magnesium sterate (+1.2° C.).

Example 6: Administration to Cynomolgus Monkeys: PolypeptideConcentration in Different Intestinal Tract Compartments and in Faeces

6.1 Polypeptide Concentration in Different Intestinal Tract Compartments

A study was conducted to assess the release profile of a compositionsimilar to that of Example 1 Batch A through regions of the intestinaltract when orally administered to Cynomolgus monkeys. The releaseprofile was assessed by analysis of polypeptide concentration in thedifferent intestinal tract compartments.

A single capsule containing 11 mini-tablets was administered orally toeach of three Cynomolgus monkeys (the monkeys are referred to as M234,M236 and M238). The mini-tablet composition varied from that of Example1 in that each mini-tablet contained an additional 1 mg of methyleneblue (dye) and a dose of 141 mg of the ICVD. 8 of the mini-tablets alsocontained 0.7 mg of isoprenaline. The methylene blue dye was for visualanalysis of the distribution of dissolved mini-tablets through thegastrointestinal (GI) tract (not discussed herein) and the isoprenalinewas for use in a study monitoring heart rate (not discussed herein).

Four hours after oral dosing, the animals were culled. Thegastrointestinal tracts were carefully removed, the different GIcompartments ligated then cut and the luminal contents and washescollected. The number of undissolved and partially dissolved minitablets were noted and these mini tablets were removed. The samples werethen homogenised and frozen until analysis. After initial centrifugationof the slurries for 5 min at 5000 rpm at 10° C., 1 ml of supernatant wasremoved from each sample and centrifuged at 13300 rpm in a microfuge atthe same temperature for 5 min. The supernatants were then centrifugedagain under the same conditions, but for 20 min, after which, they wereanalysed using a standard Humira competition ELISA (Humira is ananti-TNF-alpha antibody, also known as adalimumab). All dilutions ofsamples and Humira and the ICVD standard were prepared in PBS containing1% BSA, 0.6M NaCl, 1% human AB serum, 0.05% Tween 20 and 2× proteaseinhibitors. ICVD concentrations were interpolated from a standard curveusing a 4 parameter, non-linear curve fitting equation in GraphPadPrism. ICVD concentrations in undiluted GI tract samples and 0-4 hfaecal supernatants were derived by taking the means of the bestinterpolated data multiplied by the supernatant dilution factor.

No intact mini-tablets were found in the stomach, duodenum, jejunum orileum of either M236 or M238. In M234, 4 intact mini-tablets were foundin the stomach, 1 in the duodenum and 1 in the jejunum. No partiallydissolved mini-tablets were found in any GI tract region of any monkey.

Preparation of the slurry supernatants necessitated adding large volumesof buffer, inevitably diluting the ICVD. In FIG. 7, the expected luminalconcentrations of ICVD are presented. These were calculated, assumingthat the luminal GI tract contents have a specific gravity of 1, bymultiplying the supernatant ICVD concentrations by the fold dilution onaddition of buffer. As shown, very high ICVD (0.1→1 mM) are likely tooccur in the lumen of some monkey GI tract compartments.

ICVD was only detected in the contents of one Cynomolgus monkey stomach(M234). ICVD was also found at high concentrations in the contents ofthe ileum, caecum and upper colon of all monkeys. In addition, M234 andM238 were detected at high concentrations in the contents of the jejunum(see FIG. 7)

Finally, the % ICVD recovered was calculated, assuming the actual doseat 4 h was delivered by only mini-tablets that had dissolved. As shownin FIG. 8, between 51.5 and 74.9% of the ICVD dose was accounted for.

This study has shown that pharmaceutically active binding polypeptidecan be delivered at high concentrations to the lower GI tract ofCynomolgus monkeys. The finding that some mini-tablets remained intact 4h after dosing suggests that the dose will be delivered over a period oftime, offering the potential of prolonged exposure. If these findingsare mirrored in treatment of IBD patients when using an anti-TNF-alphabinding polypeptide, then it is reasonable to expect that theconcentrations of anti-TNF-alpha polypeptide exposed to the lower GItract will be more than adequate for effective TN F-alphaneutralisation.

6.2 Polypeptide Concentration in Faeces

A single capsule containing 11 mini-tablets was administered orally toeach of three Cynomolgus monkeys. The mini-tablet composition variedfrom that of Example 1 in that each mini-tablet contained an additional1 mg of methylene blue (dye) and 8 of the mini-tablets also contained0.7 mg of isoprenaline. The methylene blue dye was for visual analysisof the dissolution of mini-tablets in faeces and the isoprenaline wasfor use in a study monitoring heart rate (not discussed herein).

Pooled faeces from the monkeys were collected at 8, 12, 20, 24 and 36 h(no samples were collected at 16 h). No mini-tablets were found in anyof the faecal samples. These were mixed with extraction buffer (0.1%BSA, 0.6M NaCl, 0.05% Tween 20, 1× protease inhibitors, 5 mM EDTA inPBS), at 1 g faeces/4 ml buffer, then homogenised and the slurriesfrozen at −80° C. for storage before analysis. Visual examinationrevealed blue colouration of the 12 h, 20 h, 24 h and 36 h slurries.Previous in vitro experiments (not shown) have demonstrated that theincreasing methylene blue concentration upon dissolution of themini-tablets is closely correlated with ICVD concentration.

Slurries were thawed and centrifuged for 5 min at 4,000 rpm (3,200 g) toremove the bulk of particulate matter. About 1 ml of each supernatantwas transferred to Eppendorf tubes and centrifuged in a microfuge at13.5K, 10° C. for 5 min, after which supernatants were placed in newtubes and centrifuged for 20 min at 10° C. Supernatants were then usedimmediately for ICVD measurement using a Humira competition ELISA.

The ELISA OD450 readings for the different faecal supernatants are shownin FIG. 9. The data clearly show that ICVD is present in the faecessupernatant samples at all time points, with the possible exception ofthe 36 h supernatant (though there may be slight activity visible at thelowest dilution).

Interpolating these data against standard curves for ICVD using GraphPadPrism and multiplication by the dilution factor of buffer added gave theICVD concentrations in each faecal sample, using the assumptions that 1g faeces is equivalent to 1 mL liquid volume and that the polypeptide isuniformly distributed in the faeces. These are shown in FIG. 10.

Using slurry volumes (calculated on the basis of 1 g faeces=1 ml,+volume of buffer for extraction) the μg amounts of ICVD in each samplewere determined (FIG. 11)).

In summary, a sustained substantial concentration of pharmaceuticallyactive binding polypeptide was achieved through the cynomolgus monkeyintestinal tract for greater than 8 hours.

Example 7: Administration to Humans: Polypeptide Concentration at theIleal-Caecal Junction and in Faeces

7.1 Polypeptide Concentration at the Ileal-Caecal Junction

The aim of this study was to demonstrate that the pharmaceuticallyactive binding polypeptide incorporated into the composition of Example1 is delivered at high concentrations to the ileal-caecal junction inman, a major site for Crohn's and the proximal site of Crohn's lesionsin the intestine of many patients.

Four human volunteers, fitted with terminal ileostomy bags each receiveda single oral dose of 1665 mg ICVD, formulated into mini-tabs insidesize 00 capsules (9 capsules in total). In these otherwise healthyindividuals, the entire contents of the terminal ileum drains into thedetachable external bag. At each hourly time point post-dosing, thefitted bag containing the total ileal effluent was removed, frozen and anew bag was fitted. Ileostomy samples were collected in this mannerevery hour for a period of 12 hours post dosing. Following this time,ileostomy samples were collected every four hours up to 24 hours postdosing. A Pre-dosing sample (day −1) was also taken as a control. Anypartially dissolved mini-tablets observed in the bags were removed priorto analysis such that only fully soluble ICVD was analysed. The ICVD wasextracted from the ileal fluid and concentrations of active ICVD weredetermined by functional ELISA, assuming that 1 g ileal fluid isequivalent to 1 mL liquid volume.

The data revealed high concentrations of active ICVD present in theileostomy bags, in the range 200 nM up to 1 mM. In addition, highconcentrations were observed over several hours of bag changes for eachsubject (see Table 5).

TABLE 5 Hour post ICVD concentration in ileal Subject dose fluid (nM)31001 2 406350 31001 3 305560 31001 4 791 31002 2 32780 31002 3 113000031002 4 792060 31002 5 81750 31002 6 12780 31002 7 1300 31002 8 42231002 9 1410 31002 10 7520 31002 11 10080 31002 12 9210 31002 16 698031003 3 1060000 31003 4 496030 31003 5 7080 31003 8 46110 31003 9 7548031003 10 16030 31003 11 72940 31003 12 15870 31003 16 881 31004 2 12619031004 3 235 31004 4 11110 31004 5 3770 31004 6 6730 ICVD was notdetected in any of the predose (Day −1) samples from any subject.

In summary, a sustained and high concentration of pharmaceuticallyactive binding polypeptide was achieved at the ileal-caecal junction inthese human volunteers.

7.2 Polypeptide Concentration in Faeces

Healthy male subjects aged 18-45 were dosed orally with a single dose ofeither 62, 555, 1665 or 4995 mg of ICVD, using the composition detailedin Example 1. Each single dose per subject was administered between 8:30to 12:00 on day 1. Faecal samples were collected pre dose (either on day−1, or prior to dosing on day 1) and at all available times post dosingup to the morning of day 4 (the end of the study). ICVD was extractedfrom the faeces and concentrations of active ICVD were determined byfunctional ELISA, assuming that 1 g faeces is equivalent to 1 mL liquidvolume.

High concentrations in the range 180 nM to 724 μM were obtained in thefaeces of subjects (see Table 6).

TABLE 6 Faecal mg sample dose collection Pre or post Subject ID ICVD daydose [ICVD] in faeces (nM) 11001 62 −1 PRE DOSE 0 11001 1 POST DOSE 101313001 555 −1 PRE DOSE 0 13001 2 POST DOSE 1052 13003 555 −1 PRE DOSE 013003 1 POST DOSE 1938 13003 2 POST DOSE 1511 14002 1665 −1 PRE DOSE 014002 1 POST DOSE 5491 14002 2 POST DOSE 558 14004 1665 −1 PRE DOSE 014004 2 POST DOSE 27532 14006 1665 −1 PRE DOSE 0 14006 2 POST DOSE 6257915001 4995 −1 PREDOSE 0 15001 1 POST DOSE 10047 15001 2 POST DOSE 13528515001 3 POST DOSE 330 15004 4995 −1 PREDOSE 0 15004 3 POST DOSE 27315005 4995 1 PRE DOSE 0 15005 1 POST DOSE 724684 15005 2 POST DOSE258703 15005 3 POST DOSE 3536 15006 4995 −1 PRE DOSE 0 15006 1 POST DOSE57120 15006 2 POST DOSE 358 15006 2 POST DOSE 186

Anti-TNF agents that are used clinically to treat Crohn's disease, suchas adalimumab (Humira) and infliximab (Remicade), are administeredeither by intravenous infusion or subcutaneous injection. Ungar et al.(2016) Clin Gastroenterol Hepatol. 14(4):550-557 state that trough serumlevels of 56-83 nM (8-12 μg/mL) for adalimumab and 42-70 nM (6-10 μg/mL)for infliximab are required to achieve mucosal healing in 80%-90% ofpatients with IBD, and that this could be considered as a “therapeuticwindow”. These trough serum levels are also indicated in FIG. 7 inrespect of calculated luminal anti-TNF-alpha ICVD concentrations incynomolgus monkey gastrointestinal tract sections established aboveunder point 6.1.

Concentrations of anti-TNF-alpha ICVD delivered to the ileal-caecaljunction and recovered in the faeces of human volunteers during theclinical work detailed in 7.1 and 7.2 were significantly higher thanthese levels and are thus predicted to be efficacious as a treatment forCrohn's disease. This assumes that gut luminal concentrations ofanti-TNF-alpha ICVD are comparable to serum concentrations of marketedanti-TNF agents with respect to access/penetration to the gut mucosa andsub-mucosa. However, it has been demonstrated in further experimentalwork (not shown) that this anti-TNF-alpha ICVD of the invention, dosedorally in DSS colitis mice, is able to penetrate to the lamina propiawhere it is resident for several hours, despite a lack of target (TNF)engagement in mice.

Taken together with the data presented under 7.1 above, these resultsdemonstrate successful delivery of therapeutic levels of ICVD from theileal-caecal junction to the anus.

Example 8: Administration to Humans: Immunogenicity Study

Protein drugs, including therapeutic antibodies, may elicit an antibodyresponse in patients. Antibodies (of multiple Ig classes) produced inpatients that recognise epitopes of protein drugs are termed anti-drugantibodies (ADAs). The presence of ADAs can result in loss of drugefficacy/potency or adverse patient effects (van Schie et al., 2015).

A study was undertaken to assess whether sustained oral dosing in man ofthe composition of the invention elicits an ADA response. Healthy malesubjects aged 18-45 were dosed orally, three times daily, for 14 dayswith capsules containing 1665 mg (a total of 4995 mg per day) ICVD orplacebo, formulated into mini-tabs according to Example 1. Serum samplesfrom subjects were taken prior to dosing, at days 7 and 14 post-dosing,and finally at 28 days (14 days after treatment cessation). Thesesamples were analysed by Sandwich ELISA for the presence of ICVDanti-drug antibodies (ADA). This analysis revealed ADA positive sera,albeit at low titres, from 4 volunteers, two of whom received placebo.In all of these individuals ADAs were present at some level prior toICVD dosing (pre-existing ADAs).

Analysis of ICVD potency in a TNF-TNFR2 ELISA revealed that ICVDactivity against TNF-alpha was unaffected by the presence of allADA-positive human sera samples at 5%. Therefore, no evidence of ICVDneutralising ADAs was found in the sera of any volunteer at anytimepoint (see Table 7).

TABLE 7 ADA sandwich ADA ICVD Subject Active or ELISA Titre/serumneutralisa- ID placebo Sample screening dilution tion 21001 ActivePredose Negative 21001 Active Day 7 Negative 21001 Active Day 14Negative 21001 Active Day 28 Negative 21002 Active Predose Negative21002 Active Day 7 Negative 21002 Active Day 14 Negative 21002 ActiveDay 28 Negative 21003 Active Predose Negative 21003 Active Day 7Negative 21003 Active Day 14 Negative 21003 Active Day 28 Negative 21004Placebo Predose Positive 64 No 21004 Placebo Day 7 Positive 64 No 21004Placebo Day 14 Positive 64 No 21004 Placebo Day 28 Positive 64 No 21005Active Predose Positive 64 No 21005 Active Day 7 Positive 32 No 21005Active Day 14 Positive 32 No 21005 Active Day 28 Positive 32 No 21006Active Predose Negative 21006 Active Day 7 Negative 21006 Active Day 14Negative 21006 Active Day 28 Negative 21007 Active Predose Negative21007 Active Day 7 Negative 21007 Active Day 14 Negative 21007 ActiveDay 28 Negative 21008 Active Predose Positive 4 No 21008 Active Day 7Positive 4 No 21008 Active Day 14 Positive 8 No 21008 Active Day 28Positive 128 No 21009 Placebo Predose Positive 8 No 21009 Placebo Day 7Positive 8 No 21009 Placebo Day 14 Positive 16 No 21009 Placebo Day 28Positive 8 No 21010 Active Predose Negative 21010 Active Day 7 Negative21010 Active Day 14 Negative 21010 Active Day 28 Negative

Example 9: Spray Drying as an Alternative to Lyophilisation

Work was carried out to confirm that the ICVD used in the exemplifiedcomposition could be initially prepared by spray drying, instead oflyophilisation, before incorporation into the composition of theinvention.

To perform spray drying, a solution containing the ICVD is fed throughan atomiser to create a spray, which is exposed to a suitable gas streamto promote rapid evaporation. When sufficient liquid mass hasevaporated, the remaining solid material in the droplet forms anindividual particle, which is then separated from the gas stream using afilter or a cyclone.

A 3,000 mL sample of ICVD solution was used for the spray-dryingprocess. This consisted of a solution of approximately 22 mg/mL ICVD in20 mM sodium acetate (3,000 mL of 20 mM sodium acetate (MWt 82.0)equates to 4.92 g of solid. 22 mg/mL ICVD in 3000 mL equates to 66 g.Hence total solids=70.92 g of which 93.1% is ICVD). This small amount ofbuffer component was not considered to impact the spray drying processor the drug substance characteristics.

Details of the processing conditions used are provided in Table 8 below.80.4 g of spray-dried material was collected with a water content of4.4%. Hence this contained 76.86 g of dry material (consisting ofapproximately 66 g of ICVD, 4.92 g sodium acetate+traces of salts,carbohydrates and host cell proteins).

TABLE 8 Batch Size (mL) 3,000 Nozzle type 2-fluid Atomisation pressure(psig) 10 Liquid flow (g/min) 10 Drying Gas Flow (g/min) 500 Inlettemperature (° C.) 145 Outlet temperature (° C.) 60 Outlet relativehumidity (%) 15.8 Dry Powder Collected (g) 80.4 Nozzle details: SprayingSystems ¼ J Series, 1650/64 Liquid Cap/Air Cap.

Manufacture of mini-tablets from the spray-dried material followed thestandard process discussed above in respect of lyophilised material. Dueto the small quantity of spray-dried material available and hence thenumber of mini-tablets produced these had to be bulked out with placebomini-tablets of a similar size. The placebo mini-tablets were colouredbrown to distinguish them from the spray-dried tablets. On coating firstwith an HPMC base and then with a Eudragit enteric coat the browncolouring of the placebos proved an insufficient contrast and it becamedifficult to distinguish the placebos from the actives. Consequently,the enteric coating was stopped after approximately an 18% weight gainhad been achieved (against a target of 25% weight gain) to ensure thesewere not ‘over-coated’.

15 mini-tablets produced from the spray-dried material were filled intosize 00 opaque pink HPMC capsules to give a nominal dose of 185 mg ICVD,along with placebo mini-tablets for bulking.

Tests were performed on these mini-tablets and the findings are detailedin Table 9 below.

TABLE 9 Test Result Disintegration (in acid) Mini-tablets remaincomplete after 2 hours Disintegration (in phosphate buffer) Fullydisintegrated after 1 hour Content by RP-HPLC 99% Purity by RP-HPLC 92%ICVD quantitation by ELISA 125.1%   ICVD purity by SDS-PAGE 100% 

In addition, The Dynamic Dissolution Test was performed. The dissolutionprofile of the mini-tablets produced using spray-dried material wascompared to that of mini-tablets produced previously using lyophilisedmaterial (FIG. 12).

The dissolution data on the spray-dried material as expected did notmeet the optimal delay period before the coat came off once the pH ofthe medium had been raised. This was due to the thinner than target coaton these mini-tablets. For this reason, the curves in FIG. 12 for boththe lyophilised and spray dried samples have been off-set so that T=0 isthe start of release in both instances. Once the coat had come off thenthe release profile of the ICVD from the mini-tablet cores prepared byspray drying was essentially comparable to ICVD from mini-tablet coresprepared by lyophilisation.

In summary, it was concluded that the spray-dried material processedwell in the slugging and compression steps to make mini-tablets and inparticular, the resultant dry-granulated material had good flowcharacteristics. Furthermore, the drug release portion of the dynamicdissolution profile of the mini-tablets produced from spray driedmaterial was essentially comparable to that of the mini-tablets producedfrom lyophilised material.

Throughout the specification and the claims which follow, unless thecontext requires otherwise, the word ‘comprise’, and variations such as‘comprises’ and ‘comprising’, will be understood to imply the inclusionof a stated integer, step, group of integers or group of steps but notto the exclusion of any other integer, step, group of integers or groupof steps. All patents and patent applications mentioned throughout thespecification of the present invention are herein incorporated in theirentirety by reference. The invention embraces all combinations ofpreferred and more preferred groups and suitable and more suitablegroups and embodiments of groups recited above.

REFERENCES

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The invention claimed is:
 1. A solid pharmaceutical compositioncomprising a compressed core, wherein the compressed core comprises: (i)two or more pharmaceutically active binding polypeptides connected byone or more linkers, wherein the two or more pharmaceutically activebinding polypeptides are present at 40-75% by weight relative to theweight of the core, (ii) one or more compression aids wherein the one ormore compression aids is present at 20-55% by weight relative to theweight of the core, (iii) one or more disintegrants wherein the one ormore disintegrants is present at 2-6% by weight relative to the weightof the core wherein the compressed core is coated with a pH sensitiveenteric coating and wherein the two or more pharmaceutically activebinding polypeptides are immunoglobulins or immunoglobulin fragments. 2.The pharmaceutical composition according to claim 1 wherein the one ormore compression aids is selected from the list consisting of syntheticpolymers such as crospovidone, saccharides such as sucrose, glucose,lactose and fructose, sugar alcohols such as mannitol, xylitol,maltitol, erythritol, sorbitol, water-soluble polysaccharides such ascelluloses such as crystalline cellulose, microcrystalline cellulose,powdered cellulose, hydroxypropylcellulose and methyl cellulose,starches, synthetic polymers such as polyvinylpyrrolidone, sodium starchglycolate, crospovidone and inorganic compounds such as calciumcarbonate.
 3. The pharmaceutical composition according to claim 1wherein the one or more disintegrants is selected from the listconsisting of carboxymethyl cellulose, sodium carboxymethyl cellulose,croscarmellose sodium, cellulose such as low substitution degreehydroxypropylcellulose, starch such as sodium carboxymethyl starch,hydroxypropyl starch, rice starch, wheat starch, potato starch, maizestarch, partly pregelatinized starch.
 4. The pharmaceutical compositionaccording to claim 1 comprising one or more lubricants wherein the oneor more lubricants is selected from the list consisting of glycerylbehenate, a stearic acid salt such as calcium stearate, magnesiumstearate, zinc stearate, mineral oil, polyethylene glycol, sodium laurylsulfate, sodium stearyl fumarate, starch such as corn starch, potatostarch, pregelatinized starch, tapioca starch, wheat starch, stearicacid, talc, vegetable oil and zinc stearate.
 5. The pharmaceuticalcomposition according to claim 1 comprising one or more lubricantswherein the one or more lubricants is present at 0.1-2%, such as about1% by weight relative to the weight of the core.
 6. The pharmaceuticalcomposition according to claim 1 wherein the polypeptide is present atabout 50%-60% by weight relative to the weight of the core.
 7. Thepharmaceutical composition according to claim 1 wherein the pH sensitiveenteric coating has a thickness of 10-300 um.
 8. The pharmaceuticalcomposition according to claim 1 wherein the pH sensitive entericcoating comprises or consists of a pH sensitive enteric polymer coatoptionally together with one or more of a plasticiser, an anti-tackingagent and a surfactant.
 9. The pharmaceutical composition according toclaim 8 wherein the pH sensitive enteric polymer coat comprises one ormore of: methyl acrylate-methacrylic acid copolymers, cellulose acetatesuccinate, hydroxy propyl methyl cellulose phthalate, hydroxy propylmethyl cellulose acetate succinate (hypromellose acetate succinate),polyvinyl acetate phthalate (PVAP), methyl methacrylate-methacrylic acidcopolymers, sodium alginate and stearic acid.
 10. The pharmaceuticalcomposition according to claim 8 wherein the pH sensitive entericpolymer coat does not dissolve until after 2 hours or longer exposure toa pH between 0.5 and 3.5.
 11. The pharmaceutical composition accordingto claim 1 wherein the molecular weight of the polypeptide is 5-200 kD.12. The pharmaceutical composition according to claim 1 wherein thepolypeptide has an aqueous solubility of greater than 10 mg/mL, such asgreater than 30 mg/mL.
 13. The pharmaceutical composition according toclaim 1 wherein the immunoglobulin fragments are immunoglobulin chainvariable domains (ICVDs).
 14. The pharmaceutical composition accordingto claim 13 wherein the immunoglobulin chain variable domains areimmunoglobulin heavy chain variable domain.
 15. The pharmaceuticalcomposition according to claim 13 wherein the immunoglobulin chainvariable domains are immunoglobulin heavy chain variable domains from aconventional antibody (VHs) or immunoglobulin heavy chain variabledomains from a heavy chain antibody (VHHs).
 16. The pharmaceuticalcomposition according to claim 1 wherein when assayed in thePharmacopeial Dissolution Test, the pharmaceutical composition releases:(i). 10-40% by weight of the polypeptide after 30 minutes, (ii). 30-60%by weight of the polypeptide after 60 minutes and (iii). 60% by weightor greater of the polypeptide after 120 minutes.
 17. The pharmaceuticalcomposition according to claim 1 wherein when assayed in the DynamicDissolution Test: the start of release of the pharmaceutical compositionoccurs between 90 to 210 minutes, the pharmaceutical compositionreleases: (i). 10-30% by weight of the polypeptide after 60 minutes fromstart of release, (ii). 40-70% by weight of the polypeptide after 120minutes from start of release and (iii). 60% by weight or greater of thepolypeptide after 180 minutes from start of release.
 18. Thepharmaceutical composition according to claim 1 wherein when assayed inthe Pharmacopeial Enteric Coating Test, the pharmaceutical compositionreleases less than 10% by weight of the polypeptide after 2 hours. 19.The pharmaceutical composition according to claim 1 wherein the one ormore linkers are labile to one or more proteases.
 20. The pharmaceuticalcomposition according to claim 1 wherein the one or more linkers arenon-labile to one or more proteases.
 21. The pharmaceutical compositionaccording to claim 1 wherein one or more of the pharmaceutically activebinding polypeptides comprises or consists of any one of SEQ ID NOs: 1to
 28. 22. The pharmaceutical composition according to claim 21 whereinone or more of the pharmaceutically active binding polypeptidescomprises or consists of SEQ ID NO:
 28. 23. A method of delivering twoor more pharmaceutically active binding polypeptides to a region of theintestinal tract comprising orally administering a solid pharmaceuticalcomposition comprising a compressed core, wherein the compressed corecomprises: (i) the two or more pharmaceutically active bindingpolypeptides connected by one or more linkers, wherein the two or morepharmaceutically active binding polypeptides are present at 40-75% byweight relative to the weight of the core, (ii) one or more compressionaids wherein the one or more compression aids is present at 20-55% byweight relative to the weight of the core, (iii) one or moredisintegrants wherein the one or more disintegrants is present at 2-6%by weight relative to the weight of the core wherein the compressed coreis coated with a pH sensitive enteric coating and wherein the two ormore pharmaceutically active binding polypeptides are immunoglobulins orimmunoglobulin fragments.
 24. The method according to claim 23 whereinthe pH sensitive enteric coating releases polypeptide when exposed to aregion of the intestinal tract.